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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2017
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.
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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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