An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells.

An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Research Abstract Details 

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An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Abstract Text:

Bernd Bodenmiller,Lukas N Mueller,Patrick G A Pedrioli,Delphine Pflieger,Martin A ,Jimmy K Eng,Ruedi Aebersold,W Andy Tao,

Current methods for phosphoproteome analysis have several limitations. First, most methods for phosphopeptide enrichment lack the specificity to truly purify phosphopeptides. Second, fragmentation spectra of phosphopeptides, in particular those of phosphoserine and phosphothreonine containing peptides, are often dominated by the loss of the phosphate group(s) and therefore lack the information required to identify the peptide sequence and the site of phosphorylation, and third, sequence database search engines and statistical models for data validation are not optimized for the specific fragmentation properties of phosphorylated peptides. Consequently, phosphoproteomic data are characterized by large and unknown rates of false positive and false negative phosphorylation sites. Here we present an integrated chemical, mass spectrometric and computational strategy to improve the efficiency, specificity and confidence in the identification of phosphopeptides and their site(s) of phosphorylation. Phosphopeptides were isolated with high specificity through a simple derivatization procedure based on phosphoramidate chemistry. Identification of phosphopeptides, their site(s) of phosphorylation and the corresponding phosphoproteins was achieved by the optimization of the mass spectrometric data acquisition procedure, the computational tools for database searching and the data post processing. The strategy was applied to the mapping of phosphorylation sites of a purified transcription factor, dFOXO and for the global analysis of protein phosphorylation of Drosophila melanogaster Kc167 cells.

An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Publishing Authors By Initials

B Bodenmiller,LN Mueller,PG Pedrioli,D Pflieger,MA ,JK Eng,R Aebersold,WA Tao,

For similar biological sciences: biochemistry: proteomics research abstracts see: biological sciences: biochemistry: proteomics research

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An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Molecular bioSystems

VOLUME: 3

Page Numbers: 275-86

Journal Abbreviation: Mol Biosyst

ISSN: 1742-206X

DAY: 19

MONTH: 02

YEAR: 2007

An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Information

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LANGUAGE: eng

NlmUniqueID: 101251620

An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Keywords Mesh Terms:

KEYWORDS: Proteomics

MESH TERMS: methods

Chemical & Substance for Abstract: An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells. Information

Substance Name: Phosphopeptides

Registry Number: 0

Grant and Affiliation Information for An integrated chemical, mass spectrometric and computational strategy for (quantitative) phosphoproteomics: application to Drosophila melanogaster Kc167 cells.

AFFILIATION: Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland.

Country: England

AGENCY: United States NHLBI

GRANT: N01-HV-28179

ACRONYM: HV

MEDLINETA: Mol Biosyst

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