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Top-down characterization of endogenous protein complexes with native proteomics
Protein complexes exhibit great diversity in protein membership, post-translational modifications and noncovalent cofactors, enabling them to function as the actuators of many important biological processes. The exposition of these molecular features with current methods lacks either throughput or m...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5726920/ https://www.ncbi.nlm.nih.gov/pubmed/29131144 http://dx.doi.org/10.1038/nchembio.2515 |
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author | Skinner, Owen S. Haverland, Nicole A. Fornelli, Luca Melani, Rafael D. Do Vale, Luis H. F. Seckler, Henrique S. Doubleday, Peter F. Schachner, Luis F. Srzentić, Kristina Kelleher, Neil L. Compton, Philip D. |
author_facet | Skinner, Owen S. Haverland, Nicole A. Fornelli, Luca Melani, Rafael D. Do Vale, Luis H. F. Seckler, Henrique S. Doubleday, Peter F. Schachner, Luis F. Srzentić, Kristina Kelleher, Neil L. Compton, Philip D. |
author_sort | Skinner, Owen S. |
collection | PubMed |
description | Protein complexes exhibit great diversity in protein membership, post-translational modifications and noncovalent cofactors, enabling them to function as the actuators of many important biological processes. The exposition of these molecular features with current methods lacks either throughput or molecular specificity, ultimately limiting the use of protein complexes as direct analytical targets in a wide range of applications. Here, we apply native proteomics, enabled by a multistage tandem mass spectrometry approach, to characterize 125 intact endogenous complexes and 217 distinct proteoforms derived from mouse heart and human cancer cell lines in discovery mode. The native conditions preserved soluble protein–protein interactions, high-stoichiometry noncovalent cofactors, covalent modifications to cysteines, and, remarkably, superoxide ligands bound to the metal cofactor of superoxide dismutase 2. The data enable precise compositional analysis of protein complexes as they exist in the cell and demonstrate a new approach that uses mass spectrometry as a bridge to structural biology. |
format | Online Article Text |
id | pubmed-5726920 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
record_format | MEDLINE/PubMed |
spelling | pubmed-57269202018-05-13 Top-down characterization of endogenous protein complexes with native proteomics Skinner, Owen S. Haverland, Nicole A. Fornelli, Luca Melani, Rafael D. Do Vale, Luis H. F. Seckler, Henrique S. Doubleday, Peter F. Schachner, Luis F. Srzentić, Kristina Kelleher, Neil L. Compton, Philip D. Nat Chem Biol Article Protein complexes exhibit great diversity in protein membership, post-translational modifications and noncovalent cofactors, enabling them to function as the actuators of many important biological processes. The exposition of these molecular features with current methods lacks either throughput or molecular specificity, ultimately limiting the use of protein complexes as direct analytical targets in a wide range of applications. Here, we apply native proteomics, enabled by a multistage tandem mass spectrometry approach, to characterize 125 intact endogenous complexes and 217 distinct proteoforms derived from mouse heart and human cancer cell lines in discovery mode. The native conditions preserved soluble protein–protein interactions, high-stoichiometry noncovalent cofactors, covalent modifications to cysteines, and, remarkably, superoxide ligands bound to the metal cofactor of superoxide dismutase 2. The data enable precise compositional analysis of protein complexes as they exist in the cell and demonstrate a new approach that uses mass spectrometry as a bridge to structural biology. 2017-11-13 2018-01 /pmc/articles/PMC5726920/ /pubmed/29131144 http://dx.doi.org/10.1038/nchembio.2515 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Skinner, Owen S. Haverland, Nicole A. Fornelli, Luca Melani, Rafael D. Do Vale, Luis H. F. Seckler, Henrique S. Doubleday, Peter F. Schachner, Luis F. Srzentić, Kristina Kelleher, Neil L. Compton, Philip D. Top-down characterization of endogenous protein complexes with native proteomics |
title | Top-down characterization of endogenous protein complexes with native proteomics |
title_full | Top-down characterization of endogenous protein complexes with native proteomics |
title_fullStr | Top-down characterization of endogenous protein complexes with native proteomics |
title_full_unstemmed | Top-down characterization of endogenous protein complexes with native proteomics |
title_short | Top-down characterization of endogenous protein complexes with native proteomics |
title_sort | top-down characterization of endogenous protein complexes with native proteomics |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5726920/ https://www.ncbi.nlm.nih.gov/pubmed/29131144 http://dx.doi.org/10.1038/nchembio.2515 |
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