ConspectusOrganoboron compounds serve as fundamental building blocks in the construction of diverse functional molecules, ranging from pharmaceuticals to materials. The discovery of boron-related new chemical transformations serves as a pivotal driving force for developing functional substances. Nevertheless, de novo design of new reactions remains challenging, requiring both the elucidation of complex competing pathways at the molecular level and efficient navigation of high-dimensional parameter spaces of reaction conditions. Consequently, the emergence of new reaction modes still largely relies on serendipity, and progress remains slow. Recent advances in theoretical methods, computational software, and artificial intelligence have created unprecedented opportunities to accelerate new reaction discovery. This overview builds upon decades of organoboron research to demonstrate how the integration of computational and experimental studies has elucidated exceptional reactivities through precise coordination engineering, thereby establishing new, transition-metal-free synthetic methodologies based on these activation modes. Key contributions include novel activation modes for commonly used organoboron reagents such as B2pin2 and benzylic boronates, alongside unique mechanistic scenarios in B(C6F5)3 catalysis.In this Account, we begin by discussing the counterintuitive mechanisms in B(C6F5)3-catalyzed ortho-functionalization of phenols with unsaturated hydrocarbons. Our computational studies showed that the pathway does not involve a conventional Lewis acid-alkene complex. Instead, coordination of B(C6F5)3 to phenols significantly enhances the Brønsted acidity of phenolic hydroxyl groups. This enables the protonation of alkenes or alkynes and facilitates a range of reactions of ortho-functionalization to produce allylated, alkenylated, propargylic alkylated, and alkylated phenols.While diboron(4) compounds are staples in synthetic chemistry, their activation has traditionally relied on transition metals or strong bases, the latter typically leading to heterolytic B-B bond cleavage. In 2016, our comprehensive computational screening revealed that dual coordination of the boron atoms in B2pin2 by two 4-cyanopyridine molecules promotes facile homolysis, yielding pyridine-ligated boryl radicals. Further experiments confirmed the formation of resonance-stabilized radical species, while computational studies further illustrated that the related radicals can serve either as a boryl radical precursor for radical addition or as a pyridine radical for C-C coupling reactions. Such an activation mode has inspired the development of diverse radical cross-coupling reactions and gained broad adoption in boryl radical chemistry.Furthermore, we demonstrate that a computational and data-driven strategy elucidates the heterolytic cleavage of C-B bonds. By tailoring the substituents around the ate complex formed between simple boronates and alkali metal alkoxides, this activation mode generates reactive organopotassium species. Such unique reactivity unlocks diverse organometallic-type processes, including dehalogenative metalation, C-H deprotonation, and alkene addition, thereby significantly expanding the synthetic utility of these boronates beyond their conventional role as coupling partners in C-C bond formation.Perspective is given in the direction of computationally exploring uncharted chemical space to uncover exceptional reactivities in organoboron compounds. This Account is expected to offer a paradigm for advancing synthetic methodologies and efficient computational tools for a broad family of researchers.
ConspectusThe remarkable complexity of life is supported by proteins, yet their functional diversity is constrained by the limited chemical alphabet of 20 canonical amino acids. Although nature partially overcomes this restriction through nongenetically encoded processes such as post-translational modifications or cofactors, these mechanisms are often difficult to predict, control and engineer. This limitation raises a fundamental question: can we programmably "chemically edit" proteins to generate new functions on demand? To address this challenge, our laboratory has been dedicated to advancing a "protein chemical editing" toolkit by integrating synthetic chemistry with protein engineering. This framework enables precise manipulation of proteins from individual residues to entire functional domains. We pursue two complementary strategies: genetic code expansion, which introduces unnatural amino acids (UAAs) as new chemical building blocks, and directed evolution platforms, which generate programmable protein-editing enzymes capable of rewriting protein sequences.In this Account, we outline a multiscale approach for protein chemical editing, spanning atomic-level control of active sites with photocaged amino acids, refinement of catalytic pockets using noncanonical residues, covalent stabilization of protein-protein interfaces through designer electrophile warheads, and domain-level editing enabled by evolved proteases.Prospectively, through the synergistic integration of chemical design, genetic encoding, and directed evolution, protein chemical editing unlocks a new level of control over biological function. This paradigm, which merges the precision of synthetic chemistry with the complexity of living systems, fundamentally transforms our capabilities from merely observing life to actively programming it, with profound implications for biomedicine and biotechnology.
ConspectusIn 1981, the year he won the Nobel Prize, Roald Hoffmann together with Kazuyuki Tatsumi published two papers entitled "Metalloporphyrins with Unusual Geometries" that strongly influenced the state of the art in porphyrin and tetrapyrrole research at that time. The 1970s and 1980s saw the dramatic expansion of bioinorganic chemistry, using the tools of molecular coordination chemistry to model complex processes of metalloenzymes and metal cofactors. Synthetic porphyrin ligands emerged as key platforms for high-valent metal-oxo and -nitrido species, metal-metal multiple bonds and the emergence of organometallic chemistry with porphyrins as the supporting ligands. These developments were the drivers for the Hoffmann papers, which featured extended Hückel calculations to expand our then understanding of types of metalloporphyrins for which experimental evidence was just emerging, and which challenged the notion of porphyrin functioning simply as a tetradentate macrocycle with a coordinated metal ion sitting squarely in the middle.Another unquestioned assumption from that time was that porphyrin and tetrapyrrole coordination chemistry was anchored firmly in the d block of the periodic table, a not unreasonable stance given the origins of this field in heme and vitamin B12 featuring iron and cobalt. Surprisingly, the presence of group 2 element magnesium in chlorophyll had not tempted chemists to interrogate more deeply the role of main group elements in tetrapyrrole chemistry, and at the time of the Hoffmann papers, examples of s and p block elements as porphyrin complexes could be almost counted on one hand. In the nearly half century since then, as the chemistry of tetrapyrrole main group complexes has unfolded, major new examples of complexes with unusual geometries have emerged. The chemistry of the d block elements is largely governed by oxidation states and d-electron configurations while in the s and p blocks the fundamental properties of size and electronegativity dominate. Porphyrins and tetrapyrroles offer four nitrogen donors in a square-planar arrangement of fairly fixed radii, not an obvious fit for main group elements with their widely ranging sizes, electronegativities, coordination geometries and bonding types. Main group tetrapyrrole complexes are "misfits" in which the poor match between the ligand environment and the requirements of the coordinated elements stimulates unusual chemistry for both partners.In this Account, I will use the concept of metalloporphyrins with unusual geometries addressed in the Hoffmann papers to look at how tetrapyrroles bearing coordinated main group elements have extended these ideas well beyond those originally envisaged. Main group metals range from lightweight lithium to the p block heavies thallium, lead and bismuth; all are known to form porphyrin complexes, some with dramatic out-of-plane metal coordination. The classic p block elements carbon, boron, and phosphorus challenge the "metalloporphyrin" paradigm; these small, light nonmetals nevertheless exhibit a rich chemistry in a tetrapyrrole setting. The extensive range of diboron porphyrinoids feature tetrapyrroles acting as binucleating ligands, incorporating not one but two elements within the N4 coordination site. Silicon and germanium porphyrins and phthalocynanines demonstrate the interplay between redox properties of the ligand and central element. The underlying theme in this discussion will be the new concepts that can be translated into other areas of the chemical sciences.
ConspectusThe growing field of coordination supramolecular chemistry constitutes a fruitful avenue for accessing a variety of multifunctional materials with a range of applications. Their versatility is enhanced if they have the ability to encapsulate guest molecules, opening opportunities for host/guest synergies. One of the most paradigmatic categories of such assemblies is coordination supramolecular helicates, which exhibit a central cavity for the potential allocation of small species, provided that their symmetry and volumes are compatible. The presence of noncovalent interactions (NCIs) between host and guest strongly contributes to the thermodynamic stability of these edifices, sometimes giving rise to a template effect. All those features are exploited for the case of triple-stranded helicates, which are predictably obtained from reactions of metal ions that adopt an octahedral coordination geometry with ligands made of two chelating moieties sufficiently separated by a spacer. The properties of the cavity of the helicate can be tuned by adjusting the central spacer of the ligand, which in turn, may incorporate functionalities facilitating NCIs with potential guests, such as hydrogen bonds. In this manner, a collection of pyrazolylpyridine (or -quinoline) ligands (L) has given rise to a large family of (G@[M2L3])n+ species (where G represents various guests), in which the encapsulated entities are firmly held in place by [N-H···G] hydrogen bonds. These assemblies can thus be employed for the selective recognition of anions or small coordination complexes, capitalizing on the specific architecture of the ligand strands. Furthermore, they have opened a plethora of possibilities for the investigation of synergic multifunctionality. The host can be made to exhibit molecular switching behavior (for example, spin-crossover, SCO, if M = FeII) or single-ion magnet (SIM) behavior (if M = CoII) while the guest has been exploited to tune these properties or to incorporate new ones. More recently, anionic coordination complexes such as these from the series [M(ox)3]3- ("ox" being the oxalate anion and M = Fe, Cr, Al, Ru) have been efficiently trapped inside the metallo-helices. This has unveiled unprecedented phenomena resulting from encapsulation, such as the first manifestation of SIM behavior for CrIII or the enhancement of the quantum coherence of a molecular qubit when acting as the guest. This family has been expanded with the inclusion of the anilate analogues of oxalates, opening unlimited options for multiproperty explorations (such as photophysical, redox chemistry, radical generation, etc.). More recently, within this group of systems, the guest has been employed as a template to selectively assemble specific combinations of two different ligands in the form of G@[M2LxL'(3-x)]m+ heteroleptic helicates, thus leading to a further opportunity of function tunability and enhancement. In this Account, we survey this and other related types of host/guest assemblies and place them in the general context of triple-stranded supramolecular helicates while assessing their impact in fields like molecular magnetism, quantum technologies, or anion recognition.
ConspectusFour-membered nitrogen-containing heterocycles, azetidines and azetines, have recently garnered interest as attractive targets in the discovery of new compounds for pharmaceutical applications. Despite this, the full potential of these heterocycles has not yet been realized due to a dearth of general, mild synthetic methods to access them. The aza Paternò-Büchi reaction, which is the photochemical [2 + 2]-cycloaddition of imines and alkenes, provides a simple, yet powerful method for accessing azetidines. However, this transformation has historically remained limited due to inherent challenges in capturing the excited state reactivity of imines - photoexcited imines can undergo radiationless decay to the ground state through E/Z-isomerization, which precludes productive cycloaddition reactivity. For this reason, the first few decades of progress for the aza Paternò-Büchi reaction relied on cyclic imine substrates to restrict isomerization and extend the excited state lifetime of the substrate. Recently, triplet energy transfer photocatalysis has emerged as a synthetic tool for generating reactive triplet state intermediates using mild visible light irradiation and commercially available catalysts. We saw an opportunity to use triplet energy transfer to access the excited states of imines and alkenes, thus allowing for access to unprecedented classes of transformations.In this Account, we present our body of work on visible-light-mediated imine-based [2 + 2]-cycloadditions, which rely on a key design principle of energy transfer photocatalysis - careful matching of the triplet energies of the photosensitizer and substrate for selective substrate sensitization. By virtue of using visible light irradiation, this limits sensitization to activated (i.e., conjugated) alkenes and certain types of imines, and renders unactivated (i.e., unconjugated) alkenes and alkynes inaccessible. Relying on this principle, we have developed six distinct strategies (Types I-VI reactions) for accessing azetidines and azetines. These strategies differentiate intra- and intermolecular transformations and the reactivity of three distinct substrate classes: activated alkenes, unactivated alkenes, and alkynes.First, we present an intramolecular aza Paternò-Büchi reaction of acyclic oximes and hydrazones with activated alkenes (Type I). Mechanistically, this relies on selective alkene sensitization to mitigate undesired reactivity that can arise from direct excitation of the imine, meaning that this strategy is not amenable to productively engaging unactivated alkenes. To enable access to this class of substrates, we harnessed the triplet state reactivity of 2-isoxazoline-3-carboxylates in intra- (Type II) and intermolecular (Type III) aza Paternò-Büchi reactions with unactivated alkenes. We also developed a set of intermolecular reactions relying on acyclic imines and activated alkenes (Type IV), providing direct access to monocyclic azetidines for the first time under visible light conditions. Next, we present an extension of the reactivity of 2-isoxazoline-3-carboxylates with untethered (Type V) and tethered (Type VI) alkynes in intermolecular [2 + 2]-cycloadditions to generate 1- and 2-azetines. Lastly, we demonstrate the synthetic and industrial applications of our azetidine compounds: Type I products can be subjected to Ru-catalyzed oxidative β-elimination to access 1-azetines (Type VII), while Type II and III products can be synthetically modified to access nitroazetidines that have potential applications as novel energetic materials.
ConspectusGene expression of cells is a highly heterogeneous and dynamic program that changes over time in various biological processes such as embryogenesis, disease progression, and response to stimuli. Understanding the molecular mechanisms of heterogeneous and dynamic gene expression is crucial for advancing our knowledge of health and disease. The recent development of single-cell RNA sequencing (scRNA-seq) technologies has offered a great opportunity to dissect cellular heterogeneity by profiling the transcriptomes of individual cells. However, scRNA-seq captures only static snapshots of gene expression and fails to temporally resolve the RNA dynamics. Therefore, the rapid changes in transcription, the coordinated regulation of RNA synthesis and degradation rates, and the cellular interactions driving cell fate decisions remain poorly understood. In the past few years, metabolic RNA labeling-based scRNA-seq has emerged as a cutting-edge chemical tool to tackle these challenges. Nucleoside analogs are applied to label newly transcribed RNAs and distinguish them from pre-existing RNAs. This time-resolved technology unbiasedly captures the true RNA dynamics for thousands of genes in each of the individual cells, providing unprecedented insight into the regulation of heterogeneous and dynamic gene expression in diverse biological processes.In this Account, we highlight the recent advances achieved by our group and other laboratories in metabolic RNA labeling-enabled time-resolved scRNA-seq. First, we summarize the recent development of time-resolved scRNA-seq by integrating metabolic RNA labeling (e.g., 4-thioridine labeling) with various scRNA-seq platforms. We highlight our size-exclusion and locally quasi-static hydrodynamics-based Well-TEMP-seq method, which greatly improves the performance of time-resolved scRNA-seq (higher throughput, higher cell barcoding efficiency, and RNA recovery rate) and lowers the cost. Next, we extend the labeling strategy from single nucleoside labeling to double nucleoside labeling and develop scDUAL-seq The sequential (pulse-pulse) labeling by two different nucleosides in scDUAL-seq addresses the limitation of single nucleoside labeling in the simultaneous monitoring of RNA synthesis and degradation processes and accurate measurement of RNA kinetics. The ability of scDUAL-seq to discriminate between different cell states also allows the unveiling of the interplay between RNA synthesis and degradation that controls distinct RNA regulatory strategy transitions during dynamic processes. Then, we discuss the further development of in vivo metabolic RNA labeling-based scRNA-seq by our laboratory (Dyna-vivo-seq) and others, which advances the time-resolved scRNA-seq studies from cultured cells to animal models. This innovation opens new avenues to reveal single-cell RNA dynamics in living organisms. Finally, we introduce our attempts to integrate time-resolved scRNA-seq with spatial transcriptomics, adding a spatial dimension to temporal RNA dynamics. This new paradigm allows the dissection of the spatiotemporal regulation of gene expression and cell fate decisions through cell-cell interactions in the tissue microenvironment, which holds great promise for biomedical applications.Our perspectives on the current limitations of the chemical tools for single-cell RNA dynamics profiling and the future directions for improvement are also provided. We anticipate that this Account will inspire chemists to develop advanced chemical tools to profile the heterogeneous and dynamic gene expression programs and offer transformative insights into the molecular landscape of RNA dynamics in health and disease.
ConspectusThe free induction decay (FID) signal acquired in a typical NMR experiment contains information about the chemical shifts, δ, and the spin-spin coupling constants, J, of the system investigated. These two parameters, particularly the chemical shifts, are very sensitive to both intramolecular and intermolecular perturbations. As such, they are also very good probes of the structure of both the molecule and the hosting solvent/matrix. This sensitivity is exploited in natural product studies to deduce the molecular structure of newly isolated compounds from the analysis of their NMR spectra. However, for complex carbon skeletons, the interpretation of the NMR data is far from trivial; the structural information is too deeply buried within the overlapping high-order multiplets and close resonances. In these cases, it is useful to compare the experimental NMR data of the unknown substance with the ones predicted by density functional theory (DFT) based methods for hypothetical molecules. Ideally, one will discard all putative structures resulting in a disagreement with the experiments and will keep the only one exhibiting an agreement within the benchmarked accuracy of the level of theory used. Two other sources of complexity, besides the topological complexity of natural substances, may strongly affect the interpretation of the NMR spectra. One, still related with covalent compounds, is the presence of heavy atoms which brings in relativistic effects in the NMR. Even for a simple molecular structure, they turn the interpretation of the NMR spectrum into a very difficult task since empirical rules often do not allow a full elucidation of the structure; thus, such effects can be accounted for only with relativistic versions of DFT. The other source of complexity is the presence of strong noncovalent interactions of the NMR probe molecule with its environment. In these cases, the full dynamics of the solute and solvent system has to be taken into account and the structure that is responsible for the observed NMR is in fact the average bulk structure of the solute-solvent system. Then, molecular dynamics (MD) simulations have to be coupled with the DFT-NMR calculations in order to predict the NMR properties. In turn, the comparison between the calculated and experimental data can shed light on the force field (FF) parameters used in the MD simulation. Therefore, computational NMR can be used to shed light on both covalent and noncovalent structural problems: in one case, the exploration of a discrete structural space will allow one to select the correct structure of an unknown compound among several hypothesis; in the other one, it will enable the fine-tuning of classical FF parameters over a continuum range of possibilities.
ConspectusThe direct oxidation of methane, which is the main component of natural gas, shale gas, methane clathrates, and biogas, to value-added products is an economically attractive and environmentally friendly alternative to strongly endothermic methane steam reforming to synthesis gas (CO/H2). Among the different routes, the oxidative coupling of methane (OCM) to ethylene/ethane (C2-hydrocarbons) is the most promising one. A key limiting factor is insufficiently high selectivity to C2-hydrocarbons due to their overoxidation to carbon oxides (COx) at industrially relevant degrees of methane conversion. Although it is generally agreed that both selective and unselective reactions are initiated by oxygen species on the surface of catalysts, the kind, role, and origin of these species remain elusive, which hampers the tailored design of catalysts.In this Account, we summarize our recent progress in understanding how product selectivity in the OCM reaction can be tuned by controlling the type of oxygen species through catalyst composition or reaction conditions. The combination of in situ time- and temperature-resolved catalyst characterization with transient kinetic methods, i.e., temporal analysis of products (TAP) and steady-state isotopic transient kinetic analysis (SSITKA), has been proven to be effective for understanding the origin and role of oxygen species involved in selective and unselective pathways. We also present strategies for regulating the concentrations of selective and unselective oxygen species. For the Mn-M(M = Na, K, Rb, or Cs)2WO4 system, the electronegativity of the alkali metal was found to influence the ability of the catalysts to form selective oxygen species from gas-phase oxygen. The binding strength of atomic oxygen species is a key parameter for hindering the oxidation of methane to COx over Gd2O3-based catalysts. This property can be adjusted by using a metal oxide promoter. The nature and concentration of different oxygen species can also be controlled through the use of steam or an alternative oxidizing agent, N2O, and by performing the OCM reaction in a chemical looping mode, i.e., by alternating between CH4- and air-containing feeds. Using steam in the latter option enabled us to largely enhance the productivity of C2-hydrocarbons, thus making this technology more attractive for large-scale applications. The knowledge summarized in this Account is expected to present insights for further studies in the development of selective catalysts for various alkane oxidation reactions and in the optimization of reactor operation.
ConspectusFullerenes have a wide range of applications across biomedicine, electronics, and nanotechnology, yet their broader application depends on the ability to control their molecular recognition and chemical reactivity. Supramolecular host-guest chemistry offers powerful opportunities in this context by enabling selective encapsulation and modulation of reactivity within confined environments. Since their discovery, substantial efforts have been devoted to developing efficient purification protocols to obtain high-purity fullerenes (particularly higher fullerenes Cn, n > 70) from fullerene soot, thereby avoiding tedious and costly chromatographic separations. The use of molecular receptors for fullerene purification via host-guest interactions affords good selectivity, requires no specialized equipment, and enables recyclable systems through careful host design. To date, most supramolecular receptors have demonstrated differential recognition of fullerenes through distinct binding affinities, while translation of such selectivity into practical purification and, more importantly, into predictable control over reactivity remains a central challenge. Indeed, to further advance fullerene chemistry, access to isomer-pure polyfunctionalized fullerenes is essential. However, conventional functionalization methods typically yield mixtures of multiple adducts with poor regioselectivity, and chromatographic purification alone is often insufficient. An alternative strategy involves confining fullerenes within host cages that act as supramolecular masks, selectively shielding part of the fullerene surface, which has emerged as an effective approach for the direct synthesis of isomer-pure polyadducts.This Account describes our development of tetragonal prismatic nanocapsules as a unified platform for the control of fullerene recognition and reactivity. Through careful host design and detailed investigation of binding sites and binding modes, these nanocapsules enable the selective encapsulation of fullerenes with different shapes and sizes (C60, C70, C84, and fullertubes) while also providing a confined environment that governs subsequent chemical transformations, such as the regioselective functionalization of C60 and C70.In this context, supramolecular confinement not only enables selective fullerene binding but also modifies the guest's accessible surface area, thereby enabling regioselective functionalization via a supramolecular mask strategy and introducing a new level of selectivity. By systematically correlating nanocapsule structure with binding behavior and reaction outcomes, we demonstrate how cavity size matching, window accessibility, and postbinding host-guest interactions can be leveraged to achieve selective encapsulation and predictable regioselectivity in fullerene functionalization. These studies show the potential of the supramolecular control over selectivity and reactivity in highly symmetric guests.The principal remaining challenges include achieving high selectivity for specific substrates and effective discrimination among multiple competing guests through distinct, well-defined binding modes, as well as extending confinement-controlled reactivity to increasingly complex substrates. These challenges will be addressed by developing novel supramolecular containers capable of predictable recognition, enhanced selectivity, and precise control of reactivity within confined environments.
ConspectusHigh-efficiency catalytic reactions are crucial to the development of a clean and sustainable society. Thermocatalysis specializes in large-scale continuous production, but certain specific thermocatalytic processes are highly endothermic and require high operating temperatures to achieve the desirable equilibrium conversion efficiency. With the rapid development of renewable energy, electrocatalysis has drawn extensive attention because it enables green and precise chemical synthesis. Nevertheless, the electrocatalytic reaction, which undergoes a multiple-electron transfer process and suffers from inherently sluggish kinetics, faces a critical challenge for large-scale application due to its high overpotential and mass transfer limitation.Recently, the synergistic integration of thermocatalysis and electrocatalysis proposed by our group has demonstrated a series of advantages in enabling efficient catalytic reactions, which have attracted widespread research interest. The integration of thermal and electrocatalysis offers a transformative strategy that circumvents thermodynamic limitations of conventional reactions, manipulates reaction energy barriers and pathways, and thereby significantly improves the reaction rates and selectivity. Beyond these benefits, it also simplifies product separation, thereby enhancing the overall process economics. In this Account, we systematically summarize recent progress in synergistic coupling of thermocatalysis and electrocatalysis, focusing on three main strategies: (1) room-temperature thermocatalytic-electrocatalytic coupling, which circumvents traditional high reaction energy barriers via the synergy of spontaneous nonelectrochemical and electrochemical processes; (2) tandem thermocatalytic-electrocatalytic reaction, which accurately addresses the shortcomings of electrocatalytic and thermocatalytic module to break through the conversion-selectivity trade-off; and (3) an integrated thermocatalytic-electrocatalytic pathway, in which the electrochemical procedures can break the thermodynamic equilibrium of the reaction and thereby improve the overall energy efficiency. Together, these approaches provide a versatile way for constructing a high-efficiency catalytic system by revealing the design criterion of the coupling reaction process.Additionally, we discuss the key challenges and prospects in this emerging field in terms of three aspects: (i) further improving the matching degree between thermocatalysis and electrocatalysis; (ii) elucidating the mechanism of reaction activity enhancement; and (iii) trying to scale up the system for industrial-scale level production. We hope this Account will guide the development of more efficient catalytic systems in the years ahead.
ConspectusThe construction of polyfunctionalized chiral molecules represents a compelling frontier in chemical research. In this context, gold-catalyzed asymmetric alkyne transformations─achieved through the cleavage and functionalization of one or two π bonds─stand out as versatile protocols. This method features high atom economy and bond-forming efficiency, facilitating the rapid assembly of structurally diverse and complex chiral compounds. Although remarkable chemo- and regioselectivity have been established over the past decades with the development of sophisticatedly designed gold catalysts, controlling the enantioselectivity remains a challenge. This is primarily due to the outer-sphere catalytic model, which is based on the innately linear coordination geometry of gold complexes. Furthermore, achieving stereocontrol in alkyne multifunctionalization involving the cleavage of two π bonds is more complicated due to the potential catalyst dissociation during the cascade process. On the other hand, the efficiency of chiral gold catalysts has often been unpredictable, as the steric and electronic demands vary significantly across different substrates. Consequently, the development of a modular and practical strategy that leverages commercially available catalysts to achieve high enantioselectivity in alkyne functionalization remains highly desirable.In this Account, we summarize our recent efforts in developing a synergistic gold/organo catalysis strategy to address these challenges. This protocol relies on the chiral-organocatalyst-mediated asymmetric interception of two distinct classes of prochiral gold-associated intermediates: gold enolates and allylic gold species. First, leveraging the gold-catalyzed oxidative generation of gold carbenes from alkynes, we realized diverse asymmetric carbene geminal difunctionalization transformations through chiral Brønsted acid-promoted enantioselective Mannich addition of transient gold enolate species, leading to chiral dihydrofuran-3-ones and chiral α-alkoxy-β-amino ketone derivatives. Moreover, asymmetric geminal dialkylation of gold carbene species derived from unactivated internal alkynes furnishes polyfunctionalized chiral linear and cyclic ketones that incorporate quaternary stereocenters. Second, the allylic gold species, which is derived from the vinylgold intermediate via an aromatization-driven double-bond migration process, has been isolated and characterized by X-ray crystallography for the first time by our group. This key intermediate has engaged in a range of asymmetric ene-type reactions with different electrophiles in the presence of chiral quinine-derived squaramide (QN-SQA) cocatalysts, including aldol and Mannich-type addition reactions, formal Michael-type addition, stepwise [4 + 2] annulation, and divergent amination reactions. In all of these transformations, the achiral gold complexes promoted the formation of the key prochiral gold-associated intermediates under mild conditions, and the exceptionally high stereocontrol is achieved in the later interception stage facilitated by the chiral organocatalysts. A hallmark of this synergistic approach is its modularity and practicality. By an appropriate combination of achiral gold complexes with readily available chiral organocatalysts, we circumvent the lengthy search for complex chiral ligands, enabling a variety of novel asymmetric alkyne transformations by simply switching the trapping reagents with the aid of readily accessible matched chiral organocatalysts. This flexible platform not only solves long-standing stereochemical problems in alkyne functionalization but also opens new avenues for the design of distinct catalytic manifolds in asymmetric synthesis.
ConspectusOxyhalide solid-state electrolytes (SSEs) represent a strategically important subclass of halide-based materials that offer a promising solution to critical challenges in all-solid-state batteries (ASSBs), such as poor interfacial stability and mechanical fragility. By incorporating oxygen into halide frameworks, these materials preserve the wide electrochemical stability and cathode compatibility of halide SSEs while simultaneously enabling ionic conductivities exceeding 10-2 S cm-1 and enhanced thermal resilience through carefully designed oxygen incorporation routes. This unique combination makes them a frontier material class for next-generation energy storage. The precise control of oxygen content is central to optimizing oxyhalide performance. Techniques including targeted substitution reactions, nanoscale oxide additions, and the use of oxygen-rich precursors have enabled the creation of novel SSE architectures. These methods allow for meticulous defect engineering and phase purity control, which are essential for tuning bulk ionic transport and managing interfacial behavior, particularly against reactive lithium metal anodes and high-voltage cathodes operating above 4.8 V vs Li+/Li. As global efforts such as the HELENA Project and multiple academic breakthroughs converge on the development of safer and more scalable battery chemistries, oxyhalide SSEs stand out as a frontier platform with significant implications for future electric vehicles, grid storage, and aerospace energy systems.This Account provides a timely overview of the synthesis-structure-property relationships in oxyhalide SSEs. It meticulously charts the tailored chemical pathways employed in their synthesis, offering a comprehensive understanding of the design principles that govern their exceptional performance. We embark on a detailed examination of the crucial role played by diverse oxygen sources, spanning the spectrum from fundamental alkali metal oxides to sophisticated metal oxychlorides, in building the unique structural frameworks of these electrolytes. Unraveling the structure-property relationships is paramount, and this Account provides critical insights into how these oxygen sources profoundly influence the mechanisms of ion transport and ultimately dictate the overall ionic conductivity, a key metric for battery efficiency. Furthermore, we illuminate the essential characterization methodologies utilized to probe the structural, morphological, and electrochemical attributes of these fascinating materials, providing a toolkit for researchers in the field. Beyond fundamental synthesis and characterization, this Account casts a forward-looking lens onto the promising applications of oxyhalide SSEs in next-generation energy storage devices. By understanding the precise interplay between synthesis, structure, and performance, we aim to accelerate the development and implementation of safer, more energy-dense, and longer-lasting batteries for a sustainable future, impacting everything from electric vehicles to grid-scale storage.
ConspectusThe development of new methodologies for efficient and selective catalysis under mild, green conditions is critical in synthetic chemistry. While nanoconfined catalysis has emerged as a powerful way for modulating reaction pathways, existing systems lack continuously interconnected confined channels and fail to achieve precise control over the size and length of these channels, making it difficult to achieve rapid reaction at room temperature, along with difficulty in achieving ∼100% conversion and ∼100% selectivity. Learning from nature may provide a solution to this challenge. As natural catalysts, enzymes demonstrate remarkable efficiency and selectivity to certain reactions with ultralow-energy-consumption (UEC) features. Recently, a fundamental question has emerged in life sciences: how do living systems achieve UEC features in highly efficient biological processes, particularly biosynthesis? To achieve bionic UEC temperate synthesis, we propose a concept of "bioinspired nanofluidic temperate synthesis". The core innovation lies in the synergistic integration of several bioinspired design elements: (1) the precise control of channel size (interlayer spacing or pore size) to a scale comparable to molecular dimensions, which promotes the formation of "ultrafast ordered molecular fluid" and enhances reactant confinement; (2) strategic incorporation of tailored catalytic sites (e.g., Lewis and Brønsted acid, Lewis base) on the channel wall to selectively activate substrate; (3) rational incorporation of binding sites (e.g., graphitic domains, oxygen vacancies (VO)) on the channel wall to facilitate absorption of reactants. The engineered nanochannel mimics the function of an enzyme's active site and its surrounding pocket, enabling reactions to proceed along energetically favorable pathways under mild conditions. Our experimental implementation of this concept has validated its transformative potential. Stereoregulated polymerizations have been achieved using zinc porphyrin metal-organic framework membranes featuring 1D nanochannels. Utilizing 2D GO-based membranes, we have achieved high-performance flow reaction at ambient temperature, achieving ∼100% conversion and ∼100% selectivity for diverse reactions, including Knoevenagel condensation, esterification, and ring-opening reactions. Using transitional metal oxide membranes, we have achieved ∼100% conversion and ∼100% selectivity in the synthesis of acetate ester flavors and in benzylamine coupling reactions. Despite significant progress, there are still challenges to learn from the process of DNA replication resonantly driven by adenosine triphosphate releasing photons, and to develop a new reaction type, which uses low-energy photons at a specific frequency to break specific chemical bonds in a resonant manner. The rational design of active sites, fine-tuning of channel sizes, development of in situ/operando characterization methods, and application of external fields remain underexplored. This account summarizes recent progress in bioinspired nanofluidic membrane reactors and outlines future directions toward sustainable flow synthesis.
ConspectusElectrolytes underpin all electrochemical energy-storage devices by mediating ion transport, interfacial reactions, and electrochemical stability. Conventional electrolyte design has long relied on a homogeneous-solution paradigm, wherein ions are uniformly solvated and electrochemical behavior is primarily dictated by composition and bulk thermodynamic properties. Within this framework, tailoring local solvation environments has enabled important incremental improvements. However, ion transport, interfacial chemistry, and electrochemical stability remain intrinsically coupled, rendering the simultaneous optimization of multiple performance metrics fundamentally challenging.Recent experimental and computational advances have increasingly challenged the assumption of homogeneity in liquid electrolytes. Electrolytes are now recognized as structurally complex soft-matter systems that can spontaneously develop spatial heterogeneity across molecular, nanometric, and mesoscale lengths. Even in macroscopically uniform electrolytes, ions and solvent molecules may self-organize into clusters, micelles or reverse micelles, bicontinuous networks, and microemulsion-like domains. These microscopic heterostructures are not incidental: they play decisive roles in governing ion-transport pathways, interfacial reactivity, and electrochemical stability, yet have largely remained outside traditional electrolyte design frameworks.Drawing on our work over the past five years, together with related advances from the broader community, this Account introduces the concept of microheterogeneous electrolytes (MHEs), which converts electrolyte design from compositional optimization to microstructural regulation. In MHEs, spatially differentiated domains act as functional units that decouple the otherwise conflicting electrochemical requirements. Ion transport can be accelerated along percolating low-energy barrier pathways; solvent reactivity can be suppressed through confinement and topological control, and interfacial reactions are regulated via selective enrichment of active species.We first trace the historical development and thermodynamic origins of microheterogeneity (MH) in liquids, elucidating how competition between energetic and entropic contributions stabilizes nanoscale domains without macroscopic phase separation. Building on this foundation, we establish a structure-function paradigm linking solvation topology, mesoscale connectivity, and electrochemical behavior. This framework clarifies how MHEs enable fast ion transport, broaden electrochemical stability windows, and promote adaptive interphase formation under extreme conditions of temperature, voltage, and current density.To enable the rational design of MHEs, we articulate interaction-level design principles through anion and solvent. Together, these principles transform molecular interactions into programmable electrolyte architectures. We further summarize experimental and computational approaches that render these invisible structures observable and quantifiable. By redefining electrolytes as spatially organized and dynamically adaptive media, this Account establishes MHEs as a general design principle applicable to Li+, Na+, multivalent, and aqueous batteries. More broadly, it builds a conceptual bridge between soft-matter physics and electrochemical engineering, opening new opportunities for designing electrolytes capable of meeting the stringent demands of next-generation energy-storage technologies.
ConspectusThe field of covalent drug discovery has witnessed a remarkable resurgence in recent years, a trend underscored by the approval of more than 125 covalent drugs by the US FDA as of 2025, which demonstrates their immense therapeutic potential. Driven by ever-increasing computational power and vast amounts of data, deep learning (DL) is profoundly transforming numerous fields, from natural language processing to drug discovery. In the development of covalent drugs, in particular, advanced computational methods centered on data-driven approaches and artificial intelligence (AI) exhibit immense potential. The realization of this potential depends on the construction of a synergistic ecosystem. Here, we define this "ecosystem" as an integrated set of components─including (i) curated covalent-relevant databases, (ii) AI/physics-based predictive and scoring models, (iii) interoperable computational workflows spanning site identification, docking/virtual screening, and lead optimization, and (iv) closed-loop feedback that systematically incorporates experimental outcomes to update data resources and refine/validate models. This begins with the systematic collection of past experimental results to build high-quality databases. These databases, in turn, provide the foundation for developing AI-driven computational tools capable of precisely interfacing with and accelerating downstream tasks, such as molecular docking (for generating physically plausible conformations and conducting large-scale virtual screening) and lead optimization. The application of these AI tools not only guides experimental design, but the resulting key data also feed back into and enrich the databases. Furthermore, in the cutting-edge field of covalent drugs, the precise identification of "druggable" covalent sites on target proteins has emerged as another critically important downstream task.In this Account, we describe a computational and AI-driven ecosystem for structure-based covalent drug discovery and highlight our contributions to this field. By explicitly linking databases, models, workflows, and experimental feedback into a single framework, this Account moves beyond a simple inventory of individual tools to instead offer a systematic and panoramic perspective on an integrated ecosystem for covalent drug discovery, driven by data and computational engines including AI. We focus on how this ecosystem systematically addresses the challenges from covalent binding site identification to lead discovery, thereby fundamentally accelerating the development of next-generation covalent therapies. We first articulate the philosophy behind the construction and updating of covalent databases, emphasizing the necessity of high-quality data. Subsequently, we delve into a suite of cutting-edge, AI-driven computational methods, exploring the potential of deep learning in tasks such as molecular docking, covalent binding site prediction, and lead optimization. To bridge the gap between computational theory and experimental validation, we will use the discovery of potent covalent CRM1 inhibitors as a specific case study, detailing how our customized, structure-based virtual screening pipeline was utilized to achieve a seamless workflow from computational prediction to biological validation. This section is intended to offer actionable guidance for experimental researchers seeking to leverage these powerful computational tools. Finally, we highlight the limitations and potential pitfalls of this AI engine─concerns that are equally relevant when developing AI-driven covalent docking algorithms. Building on our group's recent benchmarking of AI docking methods, we objectively evaluate current performance and discuss how transformative advances such as AlphaFold3 may reshape the field.
ConspectusLanthanide-doped upconversion nanoparticles (UCNPs) represent a distinctive class of luminescent nanomaterials capable of converting low-energy near-infrared (NIR) photons into higher-energy visible or ultraviolet emission through multiphoton processes. This anti-Stokes luminescence underpins a broad spectrum of applications, including deep-tissue bioimaging, biological therapy, high-sensitivity biosensing, nanothermometry, and super-resolution microscopy. However, the upconversion brightness of UCNPs─defined as the number of photons emitted per particle per second at a given power density─remains intrinsically low, thereby constraining their widespread implementation.The weak upconversion emission brightness of UCNPs arises from a combination of intrinsic and extrinsic factors of the lanthanide ions. Intrinsically, the parity-forbidden 4f-4f electronic transitions of lanthanide ions result in low absorption cross sections and low radiative decay rates, rendering both photon absorption and emission inherently inefficient. Extrinsically, the large surface-to-volume ratio of UCNPs amplifies nonradiative energy losses through surface-related quenching, while high lanthanide dopant concentrations induce luminescence concentration quenching by nonradiative depopulation of excited states via cross-relaxation and back-energy-transfer pathways. The concentration quenching effect also precludes the possibility of reducing lanthanide interionic distances in the host lattice to accelerate energy transfer from sensitizer to emitter ions, thereby constraining the upconversion quantum yields (UCQYs). Moreover, because upconversion is a multiphoton process, the emission brightness is highly sensitive to the excitation power density, leading to a marked decrease in UCQYs under low excitation light irradiance. Additionally, uncontrolled energy migration can divert excitation energy to nonemissive sites such as crystal defects or surface quenching centers, further diminishing the luminescence efficiency. Acting individually or synergistically, these factors collectively suppress the emission brightness of UCNPs.In this Account, we review recent advances in the study of upconversion processes in lanthanide-doped nanoparticles, with a particular focus on our group's research progress over the past several years. We present a systematic framework for enhancing upconversion brightness through both intrinsic and extrinsic engineering strategies. Intrinsically, coupling UCNPs with NIR-absorbing dye sensitizers has significantly improved light harvesting, as dye molecules possess absorption cross sections approximately 4 orders of magnitude higher than those of lanthanide ions. Such sensitization has even enabled lanthanide-mediated excitation of perovskite nanocrystals under low irradiance, achieving emission enhancements exceeding 4 orders of magnitude. In parallel, plasmonic coupling accelerates radiative decay via the Purcell effect, enabling tunable emission amplification across several orders of magnitude. Extrinsically, core-ultrathick-shell architectures that leverage newly identified size-dependent lanthanide energy-transfer pathways have achieved UCQYs of up to 13%, approximately 4 times higher than their bulk counterparts. Furthermore, spatial separation of sensitizer and activator ions through core-shell nanostructuring mitigates back energy transfer and increases the concentration-quenching threshold of activator ions to as high as 50%, thereby substantially enhancing upconversion brightness by increasing the number of emitter ions. Meanwhile, lanthanide-mediated photon-avalanche processes typically introduce ultrahigh-order nonlinearities (order > 20) and yield exceptional upconversion emission intensities suitable for super-resolution imaging. Collectively, these developments deepen our understanding of nanoscale energy transfer dynamics and guide the rational design of next-generation bright UCNPs.
ConspectusDeveloping systems that can efficiently capture photon energy and convert this energy into fuels and chemicals requires understanding how to assemble molecular components with diverse functions into complete systems possessing selectivity and efficiency in directing charge carriers to catalytic reactions. There are many challenges to achieving this goal. One promising approach is the development of hybrid systems that combine semiconductor nanocrystals (NCs) for light capture and enzymes as efficient catalysts.Such biohybrid systems capitalize on the tunable electronic and optical properties of NCs while leveraging the unmatched specificity and efficiency of enzymes in catalyzing chemical reactions, thereby offering opportunities to surpass the limitations of each component alone. Here, we focus on recent progress in developing a biohybrid system that combines CdS NCs for photon capture with the enzyme nitrogenase to accomplish light-driven dinitrogen (N2) reduction to ammonia (NH3). Integrating light-harvesting materials with biological catalysts requires a deep understanding of NC properties, protein stability, and electron transfer (ET), making it an inherently multidisciplinary problem.The reduction of N2 to NH3 is a challenging reaction, with a high demand in both agriculture and industrial chemical production. This reaction is intrinsically energy intensive, due to the need to activate the N≡N triple bond. The current standard industrial approach to N2 reduction, the Haber-Bosch reaction, obtains the necessary energy input from fossil fuels, whereas biological systems capable of N2 reduction utilize the hydrolysis of ATP as their energy source. Replacing these costly, energy-intensive inputs with renewable light energy represents a critical step toward sustainable NH3 production.Recent progress has demonstrated that semiconductor CdS NCs can be coupled to the catalytic component of nitrogenase, the MoFe protein, to form a biohybrid CdS NC:MoFe protein complex, enabling light-driven N2 reduction rather than energy input from fossil fuels or ATP. This illustrates how inorganic NCs can functionally replace the natural Fe protein partner, yielding a biohybrid catalyst that enables controlled electron delivery and provides not only light-driven NH3 production but also new approaches for probing enzyme catalytic function.The CdS NC:MoFe protein biohybrid system enables light-initiated electron delivery at ambient temperature, as well as temperatures below freezing, allowing for stabilization and spectroscopic characterization of key reaction intermediates. These findings highlight how photochemical biohybrids can serve as both functional catalysts and mechanistic probes. Beyond studies of the nitrogenase mechanism, studies of the CdS NC:MoFe system reveal how variables such as NC size, electrostatic binding interactions, and sacrificial electron donors (SEDs) govern complex stability, charge transfer efficiency, and catalytic performance.In addition, studies of nitrogenase and the high activation barrier for N2 reduction are enabling investigations of new and interesting questions regarding the properties and limitations of NC biocatalysis. In this Account, we describe the key features of CdS NC:MoFe protein biohybrids and the parameters for optimal light-driven N2 reduction, and how controlling ET with light illuminates the path to new insights into the nitrogenase mechanism.
ConspectusThe direct and selective activation of chemical bonds under mild, operationally simple conditions remains a longstanding pursuit in organic synthesis. Recently, elemental σ/π-hole interactions have emerged as powerful noncovalent tools, enabling new modes of molecular activation. Despite their promise, the application of pnictogen σ/π-hole interactions in photoinduced radical processes is still at a nascent stage.In this Account, we describe our recent efforts in leveraging pnictogen σ/π-hole interactions to facilitate the generation of organic radicals under visible-light irradiation. By capitalizing on the properties of pnictogen σ/π-holes─tuned through careful selection of pnictogen elements, electron-withdrawing substituents, and pnictogen hole acceptors─we have developed general strategies for visible-light-induced radical transformations. The key element of this strategy is the use of pnictogen σ/π-hole interactions to assemble charge-transfer complexes (CTCs), which undergo visible-light-induced single-electron transfer (SET) from an electron donor to the pnictogen center. This process generates either pnictogen-centered radicals or substrate-derived radical species, thereby providing a basis for the rational design of new reagents and catalysts. The main advances can be summarized as follows:(1)We established an efficient, transition-metal-free and photocatalyst-free strategy for the generation of a broad range of radical species─including P(III)-centered, alkyl, carboranyl, fluoromethyl, difluoromethyl, trifluoromethyl, pyridyl, oxyalkyl, dn-alkyl and methylthio radicals─by using pnictogen σ-hole interactions.(2)On this basis, the scope of pnictogen-hole interaction-enabled photoreactions was further expanded by introducing N-heterocyclic nitrenium (NHN) and N-heterocyclic carbene (NHC) systems. The amphiphilic character and π-hole electron-accepting ability of NHNs promote the formation of photoactive CTCs with suitable electron donors, which, upon single-electron transfer, afford NHN-centered radicals. This approach enables a series of metal-free reductive radical transformations mediated by NHN radicals, including the activation of C-I, C-Br, and activated C-Cl bonds, as well as controlled radical polymerizations. In addition, the combination of NHNs with ligated boryl radicals allows the activation of otherwise inert alkyl chlorides, further broadening the applicability and synthetic utility of this strategy.(3)The concept of pnictogen interactions was further extended to NHC-based photocatalytic systems. NHCs, which are isoelectronic and isostructural analogues of NHNs and possess vacant pπ orbitals, can accept an electron to generate NHC radical anions. These species can act as strong reductants capable of activating a range of inert bonds, including Caryl-F, Caryl-N, Caryl-S, Caryl-Se, and Caryl-O bonds.Taken together, these advances underscore the potential of pnictogen σ/π-hole interactions in contemporary radical chemistry.
ConspectusPhotoconversion─the light-induced shift of a fluorophore's absorption and emission spectra─has attracted increasing attention across diverse fields, including single-molecule spectroscopy and super-resolution microscopy. Although blinking and photobleaching are well-established photophysical phenomena of organic dyes and have been extensively studied, photoconversion has historically received comparatively limited attention. First reported decades ago in rhodamine dyes, photoconversion has since been identified in a broad range of organic fluorophores and has become increasingly evident with advances in fluorescence imaging technologies. Unintended photoconversion can introduce spectral crosstalk and lead to misinterpretation in multicolor experiments, motivating intensified efforts to elucidate its underlying mechanisms and develop effective mitigation strategies. In this Account, we focus on the occurrence and molecular mechanisms of photoconversion in rhodamine- and cyanine-based dyes, which are widely used in commercial fluorescent probes, and discuss practical approaches to suppress its impact in fluorescence experiments. Beyond the challenge of controlling unwanted spectral shifts, we also highlight emerging opportunities to exploit photoconversion as an alternative form of "photo-switching" for biophysical applications. We first provide a historical overview of photoconversion phenomena observed in organic fluorophores, including rhodamine, cyanine, and other synthetic dye families. We then summarize the current state of research on photoconversion mechanisms. In rhodamine dyes, photobluing proceeds primarily through progressive N-dealkylation upon irradiation, which has motivated the development of synthetic strategies aimed at producing photoconversion-resistant rhodamine derivatives. Studies on cyanine dyes have revealed that photoconversion requires singlet oxygen and is strongly modulated by buffer pH. Notably, we have demonstrated by NMR spectroscopy that the photoconverted form of Cy5 is structurally identical to Cy3, a troubling observation in light of the widespread use of the Cy3-Cy5 pair as a fluorophore combination for single-molecule fluorescence resonance energy transfer (smFRET) without awareness of the underlying photoconversion. For the C2H2 excision reaction that converts Cy5 into Cy3, both intramolecular and intermolecular mechanisms have been proposed. Here, we present detailed experimental evidence supporting each photobluing pathway and offer mechanistic insights into why two distinct routes can emerge. Depending on the specific cyanine dye and experimental conditions, either pathway may dominate, or both pathways may operate concurrently within the same reaction system. We then focus on the consequences of photobluing for smFRET measurements. Building on our initial observation that even ambient light can induce Cy5 photobluing, we demonstrate that this process not only generates false donor signals but can also produce spurious FRET efficiencies. Finally, we highlight emerging applications that deliberately harness photoconversion as a new class of tools for biophysical research. By addressing its potential, we aim not only to facilitate the regulation of photoconversion in commercial dyes but also to accelerate its purposeful exploitation as a new asset in super-resolution microscopy and single-molecule spectroscopy.
ConspectusMembrane proteins perform essential functions in the complex and heterogeneous environment of the cellular membrane, where their structure and activity are profoundly influenced by the native lipid milieu. Most membrane protein structures in the Protein Data Bank (PDB) have been determined in vitro in membrane-mimetic environments such as detergents. These membrane-mimetic environments can perturb native protein conformations and may result in a loss of function, which in turn leads to misinterpretations of molecular mechanisms. Direct structural investigations of membrane proteins within native cellular membranes are crucial to understanding the undisturbed molecular mechanism. Solid-state nuclear magnetic resonance (ssNMR) spectroscopy provides a powerful platform for in situ studies of membrane proteins. However, achieving high-resolution structure determination in native membranes faces significant obstacles, including limited spectral sensitivity, background interference, and substantial uncertainties in extracting distance restraints from ssNMR spectra. These issues have long hindered complete chemical shift assignments and the determination of high-resolution structures in native environments. To address these issues, our group has established integrated in situ ssNMR methodologies, enabled by the foundational contributions of many research groups. These advances allow for complete resonance assignments, high-resolution structure determination, and residue-specific dynamics analysis of membrane proteins in native cellular membranes. This Account summarizes our contributions over the past decade to in situ ssNMR studies of membrane proteins organized around two complementary themes: methodological advances in in situ ssNMR and the elucidation of how the membrane environment influences molecular mechanisms. The discussion systematically examines (i) recent methodological progress, including selective membrane sample preparation protocols compatible with both 13C- and 1H-detected ssNMR, signal-enhancing pulse sequences, and structural computation methods and (ii) key structural and dynamic insights from in situ ssNMR, including high-resolution structure determination of diverse membrane proteins such as aquaporin Z (AqpZ), the sugar transporter BjSemiSWEET, and the channel protein MscL, alongside a mechanistic understanding of how the membrane environment regulates their functions. By demonstrating the feasibility of in situ ssNMR for determining membrane protein structures in native settings and providing unprecedented atomic-level insights into their functions, we aim to advance research on membrane-sensitive proteins and promote the broader application of in situ ssNMR methodologies.