Large-scale top-down proteomics of the human proteome: Membrane proteins, mitochondria, and senescence

132Citations
Citations of this article
142Readers
Mendeley users who have this article in their library.

This article is free to access.

Abstract

Top-down proteomics is emerging as a viable method for the routine identification of hundreds to thousands of proteins. In this work we report the largest top-down study to date, with the identification of 1,220 proteins from the transformed human cell line H1299 at a false discovery rate of 1%. Multiple separation strategies were utilized, including the focused isolation of mitochondria, resulting in significantly improved proteome coverage relative to previous work. In all, 347 mitochondrial proteins were identified, including ~50% of the mitochondrial proteome below 30 kDa and over 75% of the subunits constituting the large complexes of oxidative phosphorylation. Three hundred of the identified proteins were found to be integral membrane proteins containing between 1 and 12 transmembrane helices, requiring no specific enrichment or modified LC-MS parameters. Over 5,000 proteoforms were observed, many harboring post-translational modifications, including over a dozen proteins containing lipid anchors (some previously unknown) and many others with phosphorylation and methylation modifications. Comparison between untreated and senescent H1299 cells revealed several changes to the proteome, including the hyperphosphorylation of HMGA2. This work illustrates the burgeoning ability of top-down proteomics to characterize large numbers of intact proteoforms in a highthroughput fashion. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

Figures

References Powered by Scopus

Statistical significance for genomewide studies

7709Citations
N/AReaders
Get full text

A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids

3428Citations
N/AReaders
Get full text

Protocols to detect senescence-associated beta-galactosidase (SA-βgal) activity, a biomarker of senescent cells in culture and in vivo

1278Citations
N/AReaders
Get full text

Cited by Powered by Scopus

Progress in Top-Down Proteomics and the Analysis of Proteoforms

451Citations
N/AReaders
Get full text

Top Down proteomics: Facts and perspectives

388Citations
N/AReaders
Get full text

High-throughput proteomics

235Citations
N/AReaders
Get full text

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Cite

CITATION STYLE

APA

Catherman, A. D., Durbin, K. R., Ahl, D. R., Early, B. P., Fellers, R. T., Tran, J. C., … Kelleher, N. L. (2013). Large-scale top-down proteomics of the human proteome: Membrane proteins, mitochondria, and senescence. Molecular and Cellular Proteomics, 12(12), 3465–3473. https://doi.org/10.1074/mcp.M113.030114

Readers over time

‘13‘14‘15‘16‘17‘18‘19‘20‘21‘22‘23‘24‘2506121824

Readers' Seniority

Tooltip

PhD / Post grad / Masters / Doc 62

60%

Researcher 25

24%

Professor / Associate Prof. 15

14%

Lecturer / Post doc 2

2%

Readers' Discipline

Tooltip

Agricultural and Biological Sciences 40

38%

Biochemistry, Genetics and Molecular Bi... 30

28%

Chemistry 27

25%

Medicine and Dentistry 9

8%

Save time finding and organizing research with Mendeley

Sign up for free
0