{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T23:44:36Z","timestamp":1790379876927,"version":"4.1.0"},"reference-count":73,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T00:00:00Z","timestamp":1766016000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T00:00:00Z","timestamp":1766016000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["P20GM104316"],"award-info":[{"award-number":["P20GM104316"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:label\/>\n                    <jats:p>\n                      Yeast is a widely used model organism in biological and proteomics research. Conventional bottom\u2010up proteomic analysis of yeast cells requires disruption of the rigid cell wall to extract proteins, which is often associated with lengthy procedures, significant technical variations, and noticeable sample loss. Here, we present an \u201cin\u2010cell proteomics\u201d approach that eliminates cell lysis and digests proteins directly in the yeast cells after a rapid methanol fixation. The approach integrates all the sample processing into a single filter device, offering a simple yet highly effective and sensitive approach for yeast proteomics analysis. We applied this approach to characterize proteome dynamics in the budding yeast\n                      <jats:italic>Saccharomyces cerevisiae<\/jats:italic>\n                      during cell cycle progression and following DNA damage. With single\u2010shot LC\u2010MS, we were able to detect and quantify around 3500 yeast proteins from the in\u2010cell digests. Our study introduces a novel in\u2010cell approach for yeast proteomics analysis and presents a quantitative proteome map of yeast cell\u2010cycle progression with high temporal resolution for cell division cycle (Cdc) proteins. It also provides a comprehensive, time\u2010resolved view of proteome\u2010wide dynamics and remodeling throughout the yeast cell cycle in response to methyl methanesulfonate (MMS)\u2010induced DNA damage.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Summary<\/jats:title>\n                    <jats:p>\n                      <jats:list list-type=\"bullet\">\n                        <jats:list-item>\n                          <jats:p>\n                            Yeast proteomics studies often require detergent\u2010based and\/or mechanical disruption procedures for cell lysis and protein digestion. We reported an \u201cin\u2010cell proteomics\u201d approach that eliminates cell lysis and digests proteins directly in the yeast cells after a simple methanol fixation. The approach integrates all the sample processing into a single filter device, offering a rapid yet highly effective and sensitive approach for yeast proteomics analysis. Using this method, we were able to characterize proteome dynamics in the budding yeast\n                            <jats:italic>Saccharomyces cerevisiae<\/jats:italic>\n                            during cell cycle progression and following DNA damage.\n                          <\/jats:p>\n                        <\/jats:list-item>\n                      <\/jats:list>\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/pmic.70089","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T13:50:02Z","timestamp":1766065802000},"page":"82-93","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["In\u2010Cell Proteomics Enables High\u2010Resolution Temporal Profiling of Cell Cycle Progression and DNA Damage Response in\n                    <i>Saccharomyces cerevisiae<\/i>"],"prefix":"10.1002","volume":"26","author":[{"given":"Henry","family":"Nwaora","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry University of Delaware  Newark Delaware USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanbao","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry University of Delaware  Newark Delaware USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihao","family":"Zhuang","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry University of Delaware  Newark Delaware USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,12,18]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5330.1259"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.201800421"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/85686"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature07341"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature11835"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M113.034769"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010023\u201006739\u20105"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2020197117"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587\u2010023\u201002099\u20107"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2016.07.005"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.202300303"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11357-021-00412-3"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2020.122453"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-025-00534-0"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.3390\/proteomes13030028"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.03.026"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.TIR119.001616"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcpro.2023.100612"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msaf005"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2023.137655"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.crmeth.2024.100796"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.5c00634"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.4c05003"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.01865-24"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-025-08723-3"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1101\/2025.05.23.655823"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1101\/2025.05.20.655211"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1101\/2025.10.30.685699"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2834"},{"key":"e_1_2_8_31_1","first-page":"55","volume-title":"Cell Cycle Checkpoint Control Protocols","author":"Day A.","year":"2004"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M114.044305"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.3c02543"},{"key":"e_1_2_8_34_1","first-page":"55","volume-title":"Methods in Molecular Biology","author":"Jamur M. 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