Jacob F. Wardman, , , Feng Liu, , , Saulius Vainauskas, , , Charlotte Olagnon, , , Teresa A. Howard, , , Yuqing Tian, , , Seyed A. Nasseri, , , Rajneesh K. Bains, , , Christopher H. Taron, , and , Stephen G. Withers*,
{"title":"Reshaping of a Glycoside Hydrolase Active Site through Expression-Compensated Droplet-Based Microfluidic Screening Provides Useful Tools for Glycomics","authors":"Jacob F. Wardman, , , Feng Liu, , , Saulius Vainauskas, , , Charlotte Olagnon, , , Teresa A. Howard, , , Yuqing Tian, , , Seyed A. Nasseri, , , Rajneesh K. Bains, , , Christopher H. Taron, , and , Stephen G. Withers*, ","doi":"10.1021/acscentsci.5c01227","DOIUrl":"https://doi.org/10.1021/acscentsci.5c01227","url":null,"abstract":"<p >The glycosylation of proteins endows them with distinct biophysical properties and allows them to play fundamental roles in cellular communication. Much of our understanding of glycoproteins has derived from the ability to enzymatically manipulate glycan structures. In particular, selective cleavage of glycans from proteins simplifies the analysis of glycoproteins and the determination of structure–activity relationships. However, limited enzymatic tools are available for the study of mucin-type O-glycans. To address this, we carried out the directed evolution of a glycoside hydrolase to increase its ability to cleave the sialyl T-antigen, a ubiquitous O-glycan structure in humans. We employed ultrahigh-throughput droplet-based microfluidics to rapidly screen vast libraries of variants in pL-sized droplets, thus minimizing the quantities of complex substrate required. Furthermore, by use of fluorescent protein-fusion and ratiometric gating during droplet sorting we could account for varying expression levels and identify highly active hits that could have been overlooked due to lower expression levels. Within just two rounds of screening, we uncovered variants with 840-fold enhancements in activity and new specificities compared to those of the WT enzyme. This campaign highlights the versatility of glycoside hydrolases and provides a broadly applicable strategy to engineer enzymatic tools for glycomics through microfluidic screening.</p><p >Combining a protein expression reporter with ultrahigh-throughput droplet-based microfluidics enabled us to drastically remodel the active site of a glycoside hydrolase and engineer new activities.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1993–2005"},"PeriodicalIF":10.4,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c01227","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332082","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Wan-Chen Cindy Lee, , , Pierre-Louis Lagueux-Tremblay, , , Zongbin Jia, , and , Song Lin*,
{"title":"Regioselective Electrochemical Borylation of Oxygenated Allylic Electrophiles: Method Development and Synthetic Applications","authors":"Wan-Chen Cindy Lee, , , Pierre-Louis Lagueux-Tremblay, , , Zongbin Jia, , and , Song Lin*, ","doi":"10.1021/acscentsci.5c01074","DOIUrl":"https://doi.org/10.1021/acscentsci.5c01074","url":null,"abstract":"<p >Allylboronic esters are highly versatile intermediates in organic synthesis. In this work, we report a general and scalable strategy for the regioselective deoxygenative borylation of allylic alcohols, enals, enones, and acrylates, upgrading these abundant functional groups in feedstock chemicals and natural products into value-added borylated synthetic handles. This method achieves efficient C–O bond activation under mild electroreductive conditions, and the effective control of regioselectivity was made possible by optimizing the borylating agent and supporting electrolyte. The utility of this approach was further demonstrated in a series of telescoped synthetic sequences, enabling alcohol and carbonyl transposition, formal cross-coupling of alcohols and aldehydes, allylic amination, and vinylogous homologation. This electrosynthetic protocol offers a broadly applicable, modular route to complex allylboron compounds from simple and readily available starting materials, including terpenoid natural products.</p><p >Regioselective electrochemical borylation of allylic alcohols, enones, enals, and acrylates is reported, which enables diverse synthetic strategies to upgrade abundant feedstocks and natural products.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1959–1968"},"PeriodicalIF":10.4,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c01074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiao Huang, , , Jonathon L. Yuly, , , Peng Zhang, , , William F. DeGrado, , , Michael J. Therien, , and , David N. Beratan*,
{"title":"Design of Light Driven Hole Bifurcating Proteins","authors":"Xiao Huang, , , Jonathon L. Yuly, , , Peng Zhang, , , William F. DeGrado, , , Michael J. Therien, , and , David N. Beratan*, ","doi":"10.1021/acscentsci.5c00803","DOIUrl":"https://doi.org/10.1021/acscentsci.5c00803","url":null,"abstract":"<p >Electron bifurcation reactions divide electrons from two-electron donors into high- and low-energy pools by transporting charge on spatially separated low- and high-potential electron hopping pathways. Bifurcation delivers electrons at potentials that drive downstream reactions in photosynthesis, respiration, and biocatalysis. Recent theoretical studies have described the requirements for effective ground-state electron bifurcation. The aim of this study is to design synthetic bifurcation constructs that can be driven by light. We describe a strategy to bifurcate holes (oxidizing equivalents) efficiently with light, and we present an illustrative energy landscape that could support this design. The design focuses on the electrochemical potentials and distances between cofactors. The analysis finds that hole bifurcation may be driven efficiently with light, guiding the further development of bioinspired networks that bifurcate charge and deliver the charges with prescribed electrochemical potentials.</p><p >Bioinspired light-driven hole bifurcating networks are designed based on de novo proteins, with the aim of separating holes into spatially separated pools at different electrochemical potentials.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1911–1920"},"PeriodicalIF":10.4,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c00803","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A Conversation with Belén González-Gaya","authors":"XiaoZhi Lim, ","doi":"10.1021/acscentsci.5c01511","DOIUrl":"https://doi.org/10.1021/acscentsci.5c01511","url":null,"abstract":"<p >The environmental scientist hitched a ride on a tourist cruise to measure pollutants in Antarctica.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 9","pages":"1515–1517"},"PeriodicalIF":10.4,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c01511","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145117405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hans T. Bergal, , , Koji Kinoshita, , and , Wesley P. Wong*,
{"title":"Probing the Dynamic Strength of Biomolecular Interactions with Single-Cell Centrifugation","authors":"Hans T. Bergal, , , Koji Kinoshita, , and , Wesley P. Wong*, ","doi":"10.1021/acscentsci.5c00648","DOIUrl":"https://doi.org/10.1021/acscentsci.5c00648","url":null,"abstract":"<p >Molecular interactions between receptors and ligands govern critical biological processes, from immune surveillance and T-cell activation to tissue development. However, current techniques for studying binding avidity often sacrifice throughput or precision. We introduce a high-throughput method for quantifying molecular and cellular binding kinetics using a centrifuge force microscope (CFM)─a compact imaging system integrated into a benchtop centrifuge. The CFM performs real-time force measurements on thousands of single cells in parallel, probing receptor–ligand interactions under controlled mechanical stress. To extend these capabilities, we developed a next-generation CFM with dual-channel fluorescence imaging that enables tracking of individual cell unbinding events. To demonstrate its utility, we profiled the binding mechanics of Bispecific T-cell Engager (BiTE) molecules, immunotherapeutic proteins that facilitate T-cell targeting of cancer cells. In cell–protein assays, we quantified the avidity of T and B cells interacting with BiTE-modified surfaces, revealing receptor-specific correlations between ligand concentration and bond strength. In cell–cell assays, we characterized BiTE-mediated adhesion between Jurkat and Nalm6 cells, demonstrating a time-dependent increase in avidity. By integrating force spectroscopy with fluorescence imaging, the CFM provides a high-throughput approach for investigating the mechanochemical principles underlying receptor-mediated interactions, with broad implications for biophysical chemistry, molecular recognition, and therapeutic development.</p><p >A high-throughput Centrifuge Force Microscope enables parallel, force-based unbinding studies of molecules or cells, using fluorescence to image single-cell immunological interactions.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1946–1958"},"PeriodicalIF":10.4,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c00648","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Unveiling the Nanoconfinement Effect in CO2 Electroreduction to CH4 over Mesoporous Cu-CeO2 Nanospheres","authors":"Lei Xiong, , , Xianbiao Fu, , , Wenpu Fan, , , Jun Zhang, , , Zixuan Zheng, , , Shaojie Lu, , , Dong Wang, , , Mingze Hao, , and , Qin Yue*, ","doi":"10.1021/acscentsci.5c01035","DOIUrl":"https://doi.org/10.1021/acscentsci.5c01035","url":null,"abstract":"<p >Nanoconfinement provides a promising strategy to promote the electrochemical CO<sub>2</sub> reduction reaction (CO<sub>2</sub>RR) owing to enhanced reactant enrichment and collision. However, the nanoconfinement influence on the CH<sub>4</sub> selectivity from the CO<sub>2</sub>RR with related regulation mechanism is unclear. Herein, a series of mesoporous CeO<sub>2</sub> loaded Cu catalysts with controllable pore size (1.3–5.5 nm) are designed to modulate the CO<sub>2</sub>RR selectivity to CH<sub>4</sub>. It is found that decreasing the pore size can apparently enhance the CO<sub>2</sub>RR performance while inhibiting the HER activity. Moreover, a volcano-type relationship between the CH<sub>4</sub> selectivity and the pore diameter is observed among these catalysts, while Cu-mCeO<sub>2</sub>-3.0 (pore diameter of 3.0 nm) shows the highest CH<sub>4</sub> Faradaic efficiency (66.1 ± 2.9%). The in situ experiments and DFT calculations illustrate that a smaller pore size with stronger confinement over Cu-mCeO<sub>2</sub>-<i>x</i> can promote the adsorption and transformation of reactants (*CO, *CHO, etc.) for CH<sub>4</sub> production, but too narrow confined space (1.3 nm) will contribute to much higher intermediate coverage and promote their collision for C–C coupling to C<sub>2+</sub> products instead, thus reducing the CH<sub>4</sub> selectivity. This work provides designing insights into metal/oxide catalysts with controllable pore size to study the nanoconfinement effect on the CO<sub>2</sub>RR-to-CH<sub>4</sub> activity, which can be extended to other oxide-based catalytic reactions.</p><p >This study establishes a correlation between the CO<sub>2</sub>RR-to-CH<sub>4</sub> activity and the pore size of mesoporous Cu-CeO<sub>2</sub> catalysts, elucidating the underlying regulation mechanisms.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1902–1910"},"PeriodicalIF":10.4,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c01035","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332089","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Precisely Integrated Mesoporous Anode Enabling Fast Pseudocapacitive Sodium-Ion Storage","authors":"Shuang Li, , , Jiecheng Chen, , , Xin Miao, , , Xu Wen, , , You Zhou, , , Bingxian Chu, , , Wendi Wang, , , Yanyan Yu, , , Ziyang Guo*, , and , Kun Lan*, ","doi":"10.1021/acscentsci.5c00616","DOIUrl":"https://doi.org/10.1021/acscentsci.5c00616","url":null,"abstract":"<p >Sodium-ion batteries (SIBs) are considered potential alternatives to lithium-ion batteries (LIBs) due to the abundant resources and low sodium cost. The rational nanostructural design for anode materials plays a crucial role in SIBs. TiO<sub>2</sub>, as a common electrode material, suffers from the drawbacks of low specific surface area and poor conductivity. To overcome these limitations, we propose a strategy combining solvent evaporation-induced self-assembly and chemical oxidative polymerization to construct an ultrathin polypyrrole (PPy)-coated mesoporous TiO<sub>2</sub> microsphere (meso-TiO<sub>2</sub>@PPy) core–shell structure. The combination of the mesoporous structure and the conductive coating endows the micrometer-sized TiO<sub>2</sub> spheres with high specific surface area, excellent conductivity, and abundant sodium-ion diffusion pathways, leading to a dominant pseudocapacitance (94%) of total charge storage. Remarkably, such integration allows for a high reversible capacity of 160.6 mAh g<sup>–1</sup> at 1 A g<sup>–1</sup>, good rate performance, and stable cycling performance (capacity retention of 80.8% after 2000 cycles). Our research provides a pathway for the design of compositive anode materials for high-performance SIBs.</p><p >A type of integrated mesoporous TiO<sub>2</sub>−PPy composite is designed as an anode to guarantee high surface area, tap density, and conductivity for overall enhancement of pseudocapacitive Na<sup>+</sup> storage.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1892–1901"},"PeriodicalIF":10.4,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c00616","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A Long-Lost Coffee Bean Tastes as Good as the Best─To Understand Why, Scientists Turn to Chemistry","authors":"Marta Zaraska, ","doi":"10.1021/acscentsci.5c01454","DOIUrl":"https://doi.org/10.1021/acscentsci.5c01454","url":null,"abstract":"<p >The climate-resistant bean boasts a chemical profile similar to Arabica’s.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 9","pages":"1521–1523"},"PeriodicalIF":10.4,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c01454","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145117411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xujun Zheng, , , Chun-Yu Chiou, , , Sunay Dilara Ekim, , , Tatiana B. Kouznetsova, , , Jafer Vakil, , , Yixin Hu, , , Liel Sapir, , , Danyang Chen, , , Zi Wang, , , Michael Rubinstein, , , Jian Ping Gong, , , Nancy R. Sottos*, , and , Stephen L. Craig*,
{"title":"Tuning the Ultimate Strain of Single and Double Network Gels Through Reactive Strand Extension","authors":"Xujun Zheng, , , Chun-Yu Chiou, , , Sunay Dilara Ekim, , , Tatiana B. Kouznetsova, , , Jafer Vakil, , , Yixin Hu, , , Liel Sapir, , , Danyang Chen, , , Zi Wang, , , Michael Rubinstein, , , Jian Ping Gong, , , Nancy R. Sottos*, , and , Stephen L. Craig*, ","doi":"10.1021/acscentsci.5c00932","DOIUrl":"https://doi.org/10.1021/acscentsci.5c00932","url":null,"abstract":"<p >The stretchability (ability to be elongated) and toughness (capacity to absorb energy before breaking) of polymer network materials, such as elastomers and hydrogels, often determine their utility and lifetime. Direct correlations between the molecular behavior of polymer network components and the physical properties of the network inform the design of materials with enhanced performance, extended lifetime, and minimized waste stream. Here, we report the impact of the fused ring size in bicyclic cyclobutane mechanophores within the strands of polymer network gels. The mechanophores and their polymer strands share the same initial covalent contour length, whereas the capacity for reactive strand extension (RSE) is varied by changing the size of the ring fused to the cyclobutane from 5 to 12 carbon atoms. We observe the first evidence of covalent RSE effects in a single-network gel, and strands with greater RSE lead to gels with greater stretchability and toughness. The same qualitative correlation between molecular and macroscopic extension is also observed in DN hydrogels with mechanophores in the prestretched first network.</p><p >The strain at break of polymer network materials can be tuned by varying the molecular length hidden behind embedded cyclobutane mechanophores.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 10","pages":"1882–1891"},"PeriodicalIF":10.4,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c00932","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332086","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Noemi Jiménez-Rojo, , , Suihan Feng, , , Johannes Morstein*, , , Stefanie D. Pritzl, , , Antonino Asaro, , , Sergio López, , , Yun Xu, , , Takeshi Harayama, , , Nynke A. Vepřek, , , Christopher J. Arp, , , Martin Reynders, , , Alexander J. E. Novak, , , Evgeny Kanshin, , , Jan Lipfert, , , Beatrix Ueberheide, , , Manuel Muñiz, , , Theobald Lohmüller, , , Howard Riezman*, , and , Dirk Trauner*,
{"title":"Optical Control of Membrane Viscosity Modulates ER-to-Golgi Trafficking","authors":"Noemi Jiménez-Rojo, , , Suihan Feng, , , Johannes Morstein*, , , Stefanie D. Pritzl, , , Antonino Asaro, , , Sergio López, , , Yun Xu, , , Takeshi Harayama, , , Nynke A. Vepřek, , , Christopher J. Arp, , , Martin Reynders, , , Alexander J. E. Novak, , , Evgeny Kanshin, , , Jan Lipfert, , , Beatrix Ueberheide, , , Manuel Muñiz, , , Theobald Lohmüller, , , Howard Riezman*, , and , Dirk Trauner*, ","doi":"10.1021/acscentsci.5c00606","DOIUrl":"https://doi.org/10.1021/acscentsci.5c00606","url":null,"abstract":"<p >The lipid composition of cellular membranes is highly dynamic and undergoes continuous remodeling, affecting the biophysical properties critical to biological function. Here, we introduce an optical approach to manipulate membrane viscosity based on an exogenous synthetic fatty acid with an azobenzene photoswitch, termed <b>FAAzo4</b>. Cells rapidly incorporate <b>FAAzo4</b> into phosphatidylcholine and phosphatidylethanolamine in a concentration- and cell type-dependent manner. This generates photoswitchable PC and PE analogs, which are predominantly located in the endoplasmic reticulum. Irradiation causes a rapid photoisomerization that decreases membrane viscosity with high spatiotemporal precision. We use the resulting “PhotoCells” to study the impact of membrane viscosity on ER-to-Golgi transport and demonstrate that this two-step process has distinct membrane viscosity requirements. Our approach provides an unprecedented way of manipulating membrane biophysical properties directly in living cells and opens novel avenues to probe the effects of viscosity in a wide variety of biological processes.</p><p >PhotoCells enable the dynamic control of protein viscosity in living cells. A decrease of membrane viscosity increases the amount of protein recruited at ERES but slows down the transport to Golgi.</p>","PeriodicalId":10,"journal":{"name":"ACS Central Science","volume":"11 9","pages":"1736–1752"},"PeriodicalIF":10.4,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acscentsci.5c00606","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145117243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}