Development of an efficient, effective, and economical technology for proteome analysis

Cell Reports Methods, Journal Year: 2024, Volume and Issue: 4(6), P. 100796 - 100796

Published: June 1, 2024

We present an efficient, effective, and economical approach, named E3technology, for proteomics sample preparation. By immobilizing silica microparticles into the polytetrafluoroethylene matrix, we develop a robust membrane medium, which could serve as reliable platform to generate proteomics-friendly samples in rapid low-cost fashion. benchmark its performance using different formats demonstrate them with variety of types varied complexity, quantity, volume. Our data suggest that E3technology provides proteome-wide identification quantitation equivalent or superior many existing methods. further propose enhanced single-vessel E4technology, performs on-filter in-cell digestion minimal loss high sensitivity, enabling low-input low-cell proteomics. Lastly, utilized above technologies investigate RNA-binding proteins profile intact bacterial cell proteome.

Language: Английский

Single-nucleus proteomics identifies regulators of protein transport

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 18, 2024

The physiological response of a cell to stimulation depends on its proteome configuration. Therefore, the abundance variation regulatory proteins across unstimulated single cells can be associatively linked with their stimulation. Here we developed an approach that leverages this association individual and nuclei systematically identify potential regulators biological processes, followed by targeted validation. Specifically, applied nucleocytoplasmic protein transport in macrophages stimulated lipopolysaccharide (LPS). To end, quantified proteomes 3,412 nuclei, sampling dynamic LPS treatment, linking functional variability proteomic variability. Minutes after stimulation, correlated strongly known regulators, thus revealing impact natural cellular response. We found simple biophysical constraints, such as quantity nuclear pores, partially explain LPS-induced transport. Among many newly identified associated response, selected 16 for validation knockdown. knockdown phenotypes confirmed inferences derived from demonstrating (sub-)single-cell proteomics infer regulation. expect generalize broad applications enhance interpretability single-cell omics data.

Language: Английский

Trends in Mass Spectrometry-Based Single-Cell Proteomics

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 16, 2025

InfoMetrics Analytical ChemistryASAPArticle CiteCitationCitation and abstractCitation referencesMore citation options ShareShare onFacebookXWeChatLinkedInRedditEmailBlueskyJump toExpandCollapse ReviewMarch 16, 2025Trends in Mass Spectrometry-Based Single-Cell ProteomicsClick to copy article linkArticle link copied!Ximena Sanchez-AvilaXimena Sanchez-AvilaDepartment of Chemistry Biochemistry, Brigham Young University, Provo, Utah 84602, United StatesMore by Ximena Sanchez-AvilaView BiographyRaphaela M. de OliveiraRaphaela OliveiraDepartment Raphaela OliveiraView BiographySiqi HuangSiqi HuangDepartment Siqi HuangView BiographyChao WangChao WangDepartment Chao WangView Biographyhttps://orcid.org/0009-0008-6197-2985Ryan T. Kelly*Ryan KellyDepartment States*Email: [email protected]More Ryan KellyView Biographyhttps://orcid.org/0000-0002-3339-4443Other Access OptionsAnalytical ChemistryCite this: Anal. Chem. 2025, XXXX, XXX, XXX-XXXClick citationCitation copied!https://pubs.acs.org/doi/10.1021/acs.analchem.5c00661https://doi.org/10.1021/acs.analchem.5c00661Published March 2025 Publication History Received 28 January 2025Accepted February 2025Revised 23 2025Published online 16 2025review-article© American Chemical SocietyRequest reuse permissionsACS Publications© SocietySubjectswhat are subjects Article automatically applied from the ACS Subject Taxonomy describe scientific concepts themes article. Cells Isolation spectrometry Peptides proteins Sample preparation Note: In lieu an abstract, this is article's first page. Read To access article, please review available below. Get instant Purchase for 48 hours. Check out below using your ID or as a guest. Restore my guest Recommended through Your Institution You may have institution. institution does not content. Add change let them know you'd like include access. Through Recommend Name Loading Institutional Login Options... Change Explore subscriptions institutions Log with if you previously purchased it member benefits. hours $48.00 cart Checkout Cited By Click section linkSection copied!This has yet been cited other publications.Download PDF e-AlertsGet e-AlertsAnalytical copied!https://doi.org/10.1021/acs.analchem.5c00661Published 2025© permissionsArticle Views6Altmetric-Citations-Learn about these metrics closeArticle Views COUNTER-compliant sum full text downloads since November 2008 (both HTML) across all individuals. These regularly updated reflect usage leading up last few days.Citations number articles citing calculated Crossref daily. Find more information counts.The Altmetric Attention Score quantitative measure attention that research received online. Clicking on donut icon will load page at altmetric.com additional details score social media presence given how calculated.Recommended Articles

Language: Английский