IF 8.5

JACS Au Pub Date : 2025-05-07 DOI: 10.1021/jacsau.5c0019310.1021/jacsau.5c00193

Junling Chen*, Jiaqi Wang, Binglin Sui, Pengwei Jiang, Xumin Wang, Hongda Wang* and Feng Liang*,

{"title":"Characterizing the Membrane Assembly of ASGPR Related to Mediated Endocytosis Using TriGalNAc-Probe-Based Super-Resolution Imaging","authors":"Junling Chen*, Jiaqi Wang, Binglin Sui, Pengwei Jiang, Xumin Wang, Hongda Wang* and Feng Liang*, ","doi":"10.1021/jacsau.5c0019310.1021/jacsau.5c00193","DOIUrl":"https://doi.org/10.1021/jacsau.5c00193https://doi.org/10.1021/jacsau.5c00193","url":null,"abstract":"The asialoglycoprotein receptor (ASGPR) is a promising therapeutic target for drug delivery systems in hepatocellular carcinoma (HCC), exhibiting high affinity for specific carbohydrate residues and overexpression on malignant hepatic cells. However, their functional mechanisms remain poorly resolved at the single molecule level, hindering the rational optimization of ASGPR-targeted drug delivery systems. Here, we developed a trivalent N-acetylgalactosamine (TriGalNAc)-functionalized ligand probe leveraging high affinity to enable the nanoscale visualization of ASGPR organization and trafficking via super-resolution imaging. Fixed cell imaging revealed pronounced clustering patterns of the ASGPR on HCC membranes. In live cell experiments, we observed the distribution changes of residual ASGPR and returned ASGPR on the membrane during endocytosis, identifying protein clusters as key functional platforms for mediated ligand uptake. Additionally, comparisons with ligand probe binding under varying cell states confirmed that ASGPR aggregation degree correlates with its ligand-binding capacity. Strikingly, disruption of membrane carbohydrate cross-linking dispersed ASGPR clusters and attenuated ligand binding. These findings resolve the nanoscale assembly of ASGPR in HCC and unveil clustering-dependent ligand-binding regulation, advancing a fundamental understanding of ASGPR biology while providing new insights to refine receptor-targeted therapeutics.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2246–2256 2246–2256"},"PeriodicalIF":8.5,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/jacsau.5c00193","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144133934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Influence of a Two-Fold Ligation Pattern on Iron-Mediated Aryl-Heteroaryl Cross-Electrophile Couplings. 双重连接模式对铁介导的芳基-杂芳基交叉亲电偶联的影响。

IF 8.5

JACS Au Pub Date : 2025-05-07 eCollection Date: 2025-05-26 DOI: 10.1021/jacsau.5c00059

Faycel Djebbar, Lise-Marie Chamoreau, Guillaume Lefèvre

引用次数: 0

Developing a Cell Quenching Method to Facilitate Single Cell Mass Spectrometry Metabolomics Studies. 开发细胞猝灭方法促进单细胞质谱代谢组学研究。

IF 8.5

JACS Au Pub Date : 2025-05-06 eCollection Date: 2025-05-26 DOI: 10.1021/jacsau.5c00327

Shakya Wije Munige, Deepti Bhusal, Zongkai Peng, Dan Chen, Zhibo Yang

{"title":"Developing a Cell Quenching Method to Facilitate Single Cell Mass Spectrometry Metabolomics Studies.","authors":"Shakya Wije Munige, Deepti Bhusal, Zongkai Peng, Dan Chen, Zhibo Yang","doi":"10.1021/jacsau.5c00327","DOIUrl":"10.1021/jacsau.5c00327","url":null,"abstract":"Single-cell mass spectrometry (SCMS) has emerged as a powerful tool for analyzing metabolites in individual cells, including live cells. However, cell metabolites have a rapid turnover rate, whereas maintaining metabolites' profiles of live cells during sample transport, storage, or extended measurements can be challenging. In this study, a cell preparation method, which integrates cell washing by volatile salt solution, rapid liquid nitrogen (LN2) quenching, freeze-drying in vacuum, and freezer storage at -80 °C, to preserve cell metabolites for SCMS measurement is discussed. Experimental results revealed that LN2 quenching preserved the overall cell metabolome, whereas storage at -80 °C for 48 h slightly changed the metabolite profiles in quenched cells. However, metabolites in unquenched cells were changed regardless of low-temperature storage. The influence of omission of quenching and low-temperature storage on cell metabolites and relevant pathways were investigated. Results from this work indicate that cell quenching is necessary, but low-temperature storage time should be minimized to preserve cell metabolites. The method developed in the current work can be readily adopted by SCMS techniques with storage remaining largely unaltered, allowing for extended SCMS studies.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2379-2387"},"PeriodicalIF":8.5,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117403/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144182679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Introducing Azide into Fully Nitrated Systems: Charge Mutations toward Advanced Initiators 将叠氮化物引入完全硝化系统：向高级引发剂的电荷突变

IF 8.5

JACS Au Pub Date : 2025-05-06 DOI: 10.1021/jacsau.5c0017010.1021/jacsau.5c00170

Zhiyi Jiang, Qi Sun*, Ziqi Zhang, Ning Ding, Xuemeng Kang, Baojing Tian, Xiaoting Ren, Jinxuan He, Shenghua Li* and Siping Pang*,

{"title":"Introducing Azide into Fully Nitrated Systems: Charge Mutations toward Advanced Initiators","authors":"Zhiyi Jiang, Qi Sun*, Ziqi Zhang, Ning Ding, Xuemeng Kang, Baojing Tian, Xiaoting Ren, Jinxuan He, Shenghua Li* and Siping Pang*, ","doi":"10.1021/jacsau.5c0017010.1021/jacsau.5c00170","DOIUrl":"https://doi.org/10.1021/jacsau.5c00170https://doi.org/10.1021/jacsau.5c00170","url":null,"abstract":"Over the past century, the continuous search for advanced initiators has been a major challenge in the field of high-energy-density materials. In this study, we revealed a new strategy for constructing initiators by inducing intramolecular charge mutations, achieved by introducing a strongly charge-positive azide group into highly charge-negative fully nitrated systems. Two azide functionalized fully nitrated compounds─3-nitro-5-azide-1-trinitromethyl-1H-1,2,4-triazole (ATT) and 3-nitro-6-azide-1-trinitromethyl-1H-[1,2,4]triazolo[3,2-c][1,2,4]triazole (FTT with α and β polymorphs)─were designed and synthesized. Among these, β-FTT demonstrated a record density of 1.967 g/cm3 and detonation velocity of 9587 m/s, outperforming all previously reported organic azide compounds. Moreover, the initiation capability of β-FTT was assessed through its notably low minimum primary charge (MPC) of 30 mg, which exceeded those of representative organic initiators such as DDNP (MPC: 70 mg) and ICM-103 (MPC: 60 mg), establishing it as one of the most efficient organic initiators available. Mechanistic investigations demonstrated that charge mutations enhanced the initiating performance when compared to typical initiators and fully nitrated systems. Furthermore, high-density β-FTT demonstrated superior initiating ability relative to low-density α-FTT, challenging traditional notions regarding initiators─that density does not correlate with initiating ability. This study provides new insights into advanced initiators while emphasizing the influence of molecular charge on energy output performance.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2201–2209 2201–2209"},"PeriodicalIF":8.5,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/jacsau.5c00170","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144133887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Introducing Azide into Fully Nitrated Systems: Charge Mutations toward Advanced Initiators. 将叠氮化物引入完全硝化系统：向高级引发剂的电荷突变。

IF 8.5

JACS Au Pub Date : 2025-05-06 eCollection Date: 2025-05-26 DOI: 10.1021/jacsau.5c00170

Zhiyi Jiang, Qi Sun, Ziqi Zhang, Ning Ding, Xuemeng Kang, Baojing Tian, Xiaoting Ren, Jinxuan He, Shenghua Li, Siping Pang

{"title":"Introducing Azide into Fully Nitrated Systems: Charge Mutations toward Advanced Initiators.","authors":"Zhiyi Jiang, Qi Sun, Ziqi Zhang, Ning Ding, Xuemeng Kang, Baojing Tian, Xiaoting Ren, Jinxuan He, Shenghua Li, Siping Pang","doi":"10.1021/jacsau.5c00170","DOIUrl":"10.1021/jacsau.5c00170","url":null,"abstract":"Over the past century, the continuous search for advanced initiators has been a major challenge in the field of high-energy-density materials. In this study, we revealed a new strategy for constructing initiators by inducing intramolecular charge mutations, achieved by introducing a strongly charge-positive azide group into highly charge-negative fully nitrated systems. Two azide functionalized fully nitrated compounds3-nitro-5-azide-1-trinitromethyl-1H-1,2,4-triazole (ATT) and 3-nitro-6-azide-1-trinitromethyl-1H-[1,2,4]-triazolo-[3,2-c]-[1,2,4]-triazole (FTT with α and β polymorphs)were designed and synthesized. Among these, β-FTT demonstrated a record density of 1.967 g/cm3 and detonation velocity of 9587 m/s, outperforming all previously reported organic azide compounds. Moreover, the initiation capability of β-FTT was assessed through its notably low minimum primary charge (MPC) of 30 mg, which exceeded those of representative organic initiators such as DDNP (MPC: 70 mg) and ICM-103 (MPC: 60 mg), establishing it as one of the most efficient organic initiators available. Mechanistic investigations demonstrated that charge mutations enhanced the initiating performance when compared to typical initiators and fully nitrated systems. Furthermore, high-density β-FTT demonstrated superior initiating ability relative to low-density α-FTT, challenging traditional notions regarding initiatorsthat density does not correlate with initiating ability. This study provides new insights into advanced initiators while emphasizing the influence of molecular charge on energy output performance.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2201-2209"},"PeriodicalIF":8.5,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117400/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144183360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Developing a Cell Quenching Method to Facilitate Single Cell Mass Spectrometry Metabolomics Studies 开发细胞猝灭方法促进单细胞质谱代谢组学研究

IF 8.5

JACS Au Pub Date : 2025-05-06 DOI: 10.1021/jacsau.5c0032710.1021/jacsau.5c00327

Shakya Wije Munige, Deepti Bhusal, Zongkai Peng, Dan Chen and Zhibo Yang*,

{"title":"Developing a Cell Quenching Method to Facilitate Single Cell Mass Spectrometry Metabolomics Studies","authors":"Shakya Wije Munige, Deepti Bhusal, Zongkai Peng, Dan Chen and Zhibo Yang*, ","doi":"10.1021/jacsau.5c0032710.1021/jacsau.5c00327","DOIUrl":"https://doi.org/10.1021/jacsau.5c00327https://doi.org/10.1021/jacsau.5c00327","url":null,"abstract":"Single-cell mass spectrometry (SCMS) has emerged as a powerful tool for analyzing metabolites in individual cells, including live cells. However, cell metabolites have a rapid turnover rate, whereas maintaining metabolites’ profiles of live cells during sample transport, storage, or extended measurements can be challenging. In this study, a cell preparation method, which integrates cell washing by volatile salt solution, rapid liquid nitrogen (LN2) quenching, freeze-drying in vacuum, and freezer storage at −80 °C, to preserve cell metabolites for SCMS measurement is discussed. Experimental results revealed that LN2 quenching preserved the overall cell metabolome, whereas storage at −80 °C for 48 h slightly changed the metabolite profiles in quenched cells. However, metabolites in unquenched cells were changed regardless of low-temperature storage. The influence of omission of quenching and low-temperature storage on cell metabolites and relevant pathways were investigated. Results from this work indicate that cell quenching is necessary, but low-temperature storage time should be minimized to preserve cell metabolites. The method developed in the current work can be readily adopted by SCMS techniques with storage remaining largely unaltered, allowing for extended SCMS studies.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2379–2387 2379–2387"},"PeriodicalIF":8.5,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/jacsau.5c00327","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144133833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An Expanded View of RNA Modification with Carbohydrate-Based Metabolic Probes 以碳水化合物为基础的代谢探针对RNA修饰的扩展观点

IF 8.5

JACS Au Pub Date : 2025-05-05 DOI: 10.1021/jacsau.5c0024910.1021/jacsau.5c00249

Madoka E. Hazemi, Michael B. Geeson, Felix M. Müller, Sigitas Mikutis, Anton J. Enright* and Gonçalo J. L. Bernardes*,

{"title":"An Expanded View of RNA Modification with Carbohydrate-Based Metabolic Probes","authors":"Madoka E. Hazemi, Michael B. Geeson, Felix M. Müller, Sigitas Mikutis, Anton J. Enright* and Gonçalo J. L. Bernardes*, ","doi":"10.1021/jacsau.5c0024910.1021/jacsau.5c00249","DOIUrl":"https://doi.org/10.1021/jacsau.5c00249https://doi.org/10.1021/jacsau.5c00249","url":null,"abstract":"The discovery of glycosylated RNA (glycoRNA) revealed an unexpected association between glycans and RNA. This discovery, facilitated by the metabolic carbohydrate reporter per-O-acetylated N-azidoacetyl-mannosamine, underscores the potential for biological roles of previously unknown RNA modifications. Our study builds upon the detection of glycoRNA with other metabolic probes such as per-O-acetylated N-azidoacetyl-glucosamine, N-azidoacetyl-galactosamine, and 6-azidofucose. This broader analysis uncovered potential new glycosylated transcripts that warrant further study. However, our results also revealed unexpected resilience of the unidentified carbohydrate–RNA linkage to enzymatic cleavages by glycosidases. Therefore, we investigated nonenzymatic formation of carbohydrate–RNA linkages in vitro using per-acetylated probes. From these studies, we draw two main conclusions. First, signals arising from metabolic incorporation using acetylated carbohydrate probes represent the main source of detectable glycoRNA species. Second, sequencing reveals that the most likely candidates for carbohydrate-modified RNAs are likely tRNAs. We demonstrated that an expanded repertoire of metabolic reporters can be incorporated into glycoRNA, opening new perspectives on the nature of post-transcriptionally modified RNA.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2309–2320 2309–2320"},"PeriodicalIF":8.5,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/jacsau.5c00249","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144133855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An Expanded View of RNA Modification with Carbohydrate-Based Metabolic Probes. 以碳水化合物为基础的代谢探针对RNA修饰的扩展观点。

IF 8.5

JACS Au Pub Date : 2025-05-05 eCollection Date: 2025-05-26 DOI: 10.1021/jacsau.5c00249

Madoka E Hazemi, Michael B Geeson, Felix M Müller, Sigitas Mikutis, Anton J Enright, Gonçalo J L Bernardes

{"title":"An Expanded View of RNA Modification with Carbohydrate-Based Metabolic Probes.","authors":"Madoka E Hazemi, Michael B Geeson, Felix M Müller, Sigitas Mikutis, Anton J Enright, Gonçalo J L Bernardes","doi":"10.1021/jacsau.5c00249","DOIUrl":"10.1021/jacsau.5c00249","url":null,"abstract":"The discovery of glycosylated RNA (glycoRNA) revealed an unexpected association between glycans and RNA. This discovery, facilitated by the metabolic carbohydrate reporter per-O-acetylated N-azidoacetyl-mannosamine, underscores the potential for biological roles of previously unknown RNA modifications. Our study builds upon the detection of glycoRNA with other metabolic probes such as per-O-acetylated N-azidoacetyl-glucosamine, N-azidoacetyl-galactosamine, and 6-azidofucose. This broader analysis uncovered potential new glycosylated transcripts that warrant further study. However, our results also revealed unexpected resilience of the unidentified carbohydrate-RNA linkage to enzymatic cleavages by glycosidases. Therefore, we investigated nonenzymatic formation of carbohydrate-RNA linkages in vitro using per-acetylated probes. From these studies, we draw two main conclusions. First, signals arising from metabolic incorporation using acetylated carbohydrate probes represent the main source of detectable glycoRNA species. Second, sequencing reveals that the most likely candidates for carbohydrate-modified RNAs are likely tRNAs. We demonstrated that an expanded repertoire of metabolic reporters can be incorporated into glycoRNA, opening new perspectives on the nature of post-transcriptionally modified RNA.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2309-2320"},"PeriodicalIF":8.5,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117391/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144182678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Deep Profiling of Oocyte Aging Enabled by Simple One-Step Vial-Based Pretreatment and Single-Cell Proteomics 通过简单的一步小瓶预处理和单细胞蛋白质组学实现卵母细胞衰老的深度分析

IF 8.5

JACS Au Pub Date : 2025-05-01 DOI: 10.1021/jacsau.5c0024410.1021/jacsau.5c00244

Hui Zhang, Hailu Zhang, Chuanxi Huang, Qing Zeng, Chunyan Tian, Jing Yang, Fuchu He* and Yun Yang*,

{"title":"Deep Profiling of Oocyte Aging Enabled by Simple One-Step Vial-Based Pretreatment and Single-Cell Proteomics","authors":"Hui Zhang, Hailu Zhang, Chuanxi Huang, Qing Zeng, Chunyan Tian, Jing Yang, Fuchu He* and Yun Yang*, ","doi":"10.1021/jacsau.5c0024410.1021/jacsau.5c00244","DOIUrl":"https://doi.org/10.1021/jacsau.5c00244https://doi.org/10.1021/jacsau.5c00244","url":null,"abstract":"Single-cell proteomics is a pivotal technology for studying cellular phenotypes, offering unparalleled insights into cellular heterogeneity and dynamic functions. Technical improvement in mass spectrometry instruments and sample preparation has made proteomics profiling of single mouse oocytes or early embryos feasible in recent years. Yet, developing a simple and robust sample preparation method to enable deep proteomics profiling of single germline cells remains a significant challenge. Herein, we developed a simple one-step vial-based pretreatment (SOViP) for deep label-free single-cell proteomics of germline cells. SOViP integrates all sample preparation procedures into a single step in autosampler vials, yet it is highly efficient and high-throughput in comparison to reported multistep methods. SOViP can be finished within ∼2 h with hands-on time limited to merely a few minutes. On average, over 6500 protein groups can be quantified from a single mouse oocyte using SOViP. In total, 6983 protein groups were identified from single mouse oocytes across an entire reproductive lifespan, offering a valuable proteomics resource for oocyte aging. Unique molecular characteristics of oocytes at different ages were revealed, and a classifier consisting of ten proteins demonstrated accurate age-group classification and fertility-level prediction. Although demonstrated using mouse oocytes in this study, SOViP is adaptable to rare cell samples and other large cells including follicles and preimplantation embryo cells, among others.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2321–2333 2321–2333"},"PeriodicalIF":8.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/jacsau.5c00244","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144133944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Deep Profiling of Oocyte Aging Enabled by Simple One-Step Vial-Based Pretreatment and Single-Cell Proteomics. 通过简单的一步小瓶预处理和单细胞蛋白质组学实现卵母细胞衰老的深度分析。

IF 8.5

JACS Au Pub Date : 2025-05-01 eCollection Date: 2025-05-26 DOI: 10.1021/jacsau.5c00244

Hui Zhang, Hailu Zhang, Chuanxi Huang, Qing Zeng, Chunyan Tian, Jing Yang, Fuchu He, Yun Yang

{"title":"Deep Profiling of Oocyte Aging Enabled by Simple One-Step Vial-Based Pretreatment and Single-Cell Proteomics.","authors":"Hui Zhang, Hailu Zhang, Chuanxi Huang, Qing Zeng, Chunyan Tian, Jing Yang, Fuchu He, Yun Yang","doi":"10.1021/jacsau.5c00244","DOIUrl":"10.1021/jacsau.5c00244","url":null,"abstract":"Single-cell proteomics is a pivotal technology for studying cellular phenotypes, offering unparalleled insights into cellular heterogeneity and dynamic functions. Technical improvement in mass spectrometry instruments and sample preparation has made proteomics profiling of single mouse oocytes or early embryos feasible in recent years. Yet, developing a simple and robust sample preparation method to enable deep proteomics profiling of single germline cells remains a significant challenge. Herein, we developed a simple one-step vial-based pretreatment (SOViP) for deep label-free single-cell proteomics of germline cells. SOViP integrates all sample preparation procedures into a single step in autosampler vials, yet it is highly efficient and high-throughput in comparison to reported multistep methods. SOViP can be finished within ∼2 h with hands-on time limited to merely a few minutes. On average, over 6500 protein groups can be quantified from a single mouse oocyte using SOViP. In total, 6983 protein groups were identified from single mouse oocytes across an entire reproductive lifespan, offering a valuable proteomics resource for oocyte aging. Unique molecular characteristics of oocytes at different ages were revealed, and a classifier consisting of ten proteins demonstrated accurate age-group classification and fertility-level prediction. Although demonstrated using mouse oocytes in this study, SOViP is adaptable to rare cell samples and other large cells including follicles and preimplantation embryo cells, among others.","PeriodicalId":94060,"journal":{"name":"JACS Au","volume":"5 5","pages":"2321-2333"},"PeriodicalIF":8.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12117414/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144183209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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