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High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms
Single-cell proteomics has emerged as a powerful method to characterize cellular phenotypic heterogeneity and the cell-specific functional networks underlying biological processes. However, significant challenges remain in single-cell proteomics for the analysis of proteoforms arising from genetic m...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10175728/ https://www.ncbi.nlm.nih.gov/pubmed/37126723 http://dx.doi.org/10.1073/pnas.2222081120 |
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author | Melby, Jake A. Brown, Kyle A. Gregorich, Zachery R. Roberts, David S. Chapman, Emily A. Ehlers, Lauren E. Gao, Zhan Larson, Eli J. Jin, Yutong Lopez, Justin R. Hartung, Jared Zhu, Yanlong McIlwain, Sean J. Wang, Daojing Guo, Wei Diffee, Gary M. Ge, Ying |
author_facet | Melby, Jake A. Brown, Kyle A. Gregorich, Zachery R. Roberts, David S. Chapman, Emily A. Ehlers, Lauren E. Gao, Zhan Larson, Eli J. Jin, Yutong Lopez, Justin R. Hartung, Jared Zhu, Yanlong McIlwain, Sean J. Wang, Daojing Guo, Wei Diffee, Gary M. Ge, Ying |
author_sort | Melby, Jake A. |
collection | PubMed |
description | Single-cell proteomics has emerged as a powerful method to characterize cellular phenotypic heterogeneity and the cell-specific functional networks underlying biological processes. However, significant challenges remain in single-cell proteomics for the analysis of proteoforms arising from genetic mutations, alternative splicing, and post-translational modifications. Herein, we have developed a highly sensitive functionally integrated top–down proteomics method for the comprehensive analysis of proteoforms from single cells. We applied this method to single muscle fibers (SMFs) to resolve their heterogeneous functional and proteomic properties at the single-cell level. Notably, we have detected single-cell heterogeneity in large proteoforms (>200 kDa) from the SMFs. Using SMFs obtained from three functionally distinct muscles, we found fiber-to-fiber heterogeneity among the sarcomeric proteoforms which can be related to the functional heterogeneity. Importantly, we detected multiple isoforms of myosin heavy chain (~223 kDa), a motor protein that drives muscle contraction, with high reproducibility to enable the classification of individual fiber types. This study reveals single muscle cell heterogeneity in large proteoforms and establishes a direct relationship between sarcomeric proteoforms and muscle fiber types, highlighting the potential of top–down proteomics for uncovering the molecular underpinnings of cell-to-cell variation in complex systems. |
format | Online Article Text |
id | pubmed-10175728 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-101757282023-11-01 High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms Melby, Jake A. Brown, Kyle A. Gregorich, Zachery R. Roberts, David S. Chapman, Emily A. Ehlers, Lauren E. Gao, Zhan Larson, Eli J. Jin, Yutong Lopez, Justin R. Hartung, Jared Zhu, Yanlong McIlwain, Sean J. Wang, Daojing Guo, Wei Diffee, Gary M. Ge, Ying Proc Natl Acad Sci U S A Biological Sciences Single-cell proteomics has emerged as a powerful method to characterize cellular phenotypic heterogeneity and the cell-specific functional networks underlying biological processes. However, significant challenges remain in single-cell proteomics for the analysis of proteoforms arising from genetic mutations, alternative splicing, and post-translational modifications. Herein, we have developed a highly sensitive functionally integrated top–down proteomics method for the comprehensive analysis of proteoforms from single cells. We applied this method to single muscle fibers (SMFs) to resolve their heterogeneous functional and proteomic properties at the single-cell level. Notably, we have detected single-cell heterogeneity in large proteoforms (>200 kDa) from the SMFs. Using SMFs obtained from three functionally distinct muscles, we found fiber-to-fiber heterogeneity among the sarcomeric proteoforms which can be related to the functional heterogeneity. Importantly, we detected multiple isoforms of myosin heavy chain (~223 kDa), a motor protein that drives muscle contraction, with high reproducibility to enable the classification of individual fiber types. This study reveals single muscle cell heterogeneity in large proteoforms and establishes a direct relationship between sarcomeric proteoforms and muscle fiber types, highlighting the potential of top–down proteomics for uncovering the molecular underpinnings of cell-to-cell variation in complex systems. National Academy of Sciences 2023-05-01 2023-05-09 /pmc/articles/PMC10175728/ /pubmed/37126723 http://dx.doi.org/10.1073/pnas.2222081120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Biological Sciences Melby, Jake A. Brown, Kyle A. Gregorich, Zachery R. Roberts, David S. Chapman, Emily A. Ehlers, Lauren E. Gao, Zhan Larson, Eli J. Jin, Yutong Lopez, Justin R. Hartung, Jared Zhu, Yanlong McIlwain, Sean J. Wang, Daojing Guo, Wei Diffee, Gary M. Ge, Ying High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
title | High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
title_full | High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
title_fullStr | High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
title_full_unstemmed | High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
title_short | High sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
title_sort | high sensitivity top–down proteomics captures single muscle cell heterogeneity in large proteoforms |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10175728/ https://www.ncbi.nlm.nih.gov/pubmed/37126723 http://dx.doi.org/10.1073/pnas.2222081120 |
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