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PMID: 15302935 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Large-scale characterization of HeLa cell nuclear phosphoproteins.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 33 ·2004-08-17 ·Pages 12130-5

Beausoleil SA, Jedrychowski M, Schwartz D, Elias JE, Villén J, Li J, Cohn MA, Cantley LC, Gygi SP

Abstract

Determining the site of a regulatory phosphorylation event is often essential for elucidating specific kinase-substrate relationships, providing a handle for understanding essential signaling pathways and ultimately allowing insights into numerous disease pathologies. Despite intense research efforts to elucidate mechanisms of protein phosphorylation regulation, efficient, large-scale identification and characterization of phosphorylation sites remains an unsolved problem. In this report we describe an application of existing technology for the isolation and identification of phosphorylation sites. By using a strategy based on strong cation exchange chromatography, phosphopeptides were enriched from the nuclear fraction of HeLa cell lysate. From 967 proteins, 2,002 phosphorylation sites were determined by tandem MS. This unprecedented large collection of sites permitted a detailed accounting of known and unknown kinase motifs and substrates.

MeSH Terms
Amino Acid Sequence Binding Sites Cell Nucleus/metabolism Chromatography, Ion Exchange Computational Biology HeLa Cells Humans Mass Spectrometry Molecular Sequence Data Nuclear Proteins/chemistry,genetics,isolation & purification,metabolism Phosphoproteins/chemistry,genetics,isolation & purification,metabolism Phosphorylation Serine/chemistry Threonine/chemistry Trypsin
Chemicals
Nuclear Proteins Phosphoproteins Threonine Serine Trypsin
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Beausoleil Sean A
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Jedrychowski Mark
Schwartz Daniel
Elias Joshua E
Villén Judit
Li Jiaxu
Cohn Martin A
Cantley Lewis C
Gygi Steven P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0404720101
Published
2004-08-17
Epub
2004-00-09
Pages
12130-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC514446
Subset
IM
Grants
NHGRI NIH HHS · HG00041 · United States
NIGMS NIH HHS · GMS6203 · United States
NHGRI NIH HHS · T32 HG000041 · United States
NHGRI NIH HHS · K22 HG000041 · United States
NIGMS NIH HHS · R01 GM067945 · United States
NIGMS NIH HHS · GM67945 · United States
NIGMS NIH HHS · R01 GM056203 · United States
External Links
PubMed source DOI: 10.1073/pnas.0404720101 PMC Full Text View journal details
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