Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics.

Blagoy Blagoev, S.E. Ong, Irina Kratchmarova, Matthias Mann

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

To study the global dynamics of phosphotyrosine-based signaling events in early growth factor stimulation, we developed a mass spectrometric method that converts temporal changes to differences in peptide isotopic abundance. The proteomes of three cell populations were metabolically encoded with different stable isotopic forms of arginine. Each population was stimulated by epidermal growth factor for a different length of time, and tyrosine-phosphorylated proteins and closely associated binders were affinity purified. Arginine-containing peptides occurred in three forms, which were quantified; we then combined two experiments to generate five-point dynamic profiles. We identified 81 signaling proteins, including virtually all known epidermal growth factor receptor substrates, 31 novel effectors and the time course of their activation upon epidermal growth factor stimulation. Global activation profiles provide an informative perspective on cell signaling and will be crucial to modeling signaling networks in a systems biology approach.
OriginalsprogEngelsk
TidsskriftNature Biotechnology
Vol/bind22
Sider (fra-til)1139-1145
ISSN1087-0156
DOI
StatusUdgivet - 2004

Fingeraftryk

Phosphotyrosine
Arginine
Proteins
Epidermal Growth Factor
Peptides
Chemical activation
Cell signaling
Systems Biology
Proteome
Epidermal Growth Factor Receptor
Population
Binders
Tyrosine
Intercellular Signaling Peptides and Proteins
Cells
Substrates
Proteomics
Experiments

Citer dette

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abstract = "To study the global dynamics of phosphotyrosine-based signaling events in early growth factor stimulation, we developed a mass spectrometric method that converts temporal changes to differences in peptide isotopic abundance. The proteomes of three cell populations were metabolically encoded with different stable isotopic forms of arginine. Each population was stimulated by epidermal growth factor for a different length of time, and tyrosine-phosphorylated proteins and closely associated binders were affinity purified. Arginine-containing peptides occurred in three forms, which were quantified; we then combined two experiments to generate five-point dynamic profiles. We identified 81 signaling proteins, including virtually all known epidermal growth factor receptor substrates, 31 novel effectors and the time course of their activation upon epidermal growth factor stimulation. Global activation profiles provide an informative perspective on cell signaling and will be crucial to modeling signaling networks in a systems biology approach.",
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Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics. / Blagoev, Blagoy; Ong, S.E.; Kratchmarova, Irina; Mann, Matthias.

I: Nature Biotechnology, Bind 22, 2004, s. 1139-1145.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics.

AU - Blagoev, Blagoy

AU - Ong, S.E.

AU - Kratchmarova, Irina

AU - Mann, Matthias

PY - 2004

Y1 - 2004

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AB - To study the global dynamics of phosphotyrosine-based signaling events in early growth factor stimulation, we developed a mass spectrometric method that converts temporal changes to differences in peptide isotopic abundance. The proteomes of three cell populations were metabolically encoded with different stable isotopic forms of arginine. Each population was stimulated by epidermal growth factor for a different length of time, and tyrosine-phosphorylated proteins and closely associated binders were affinity purified. Arginine-containing peptides occurred in three forms, which were quantified; we then combined two experiments to generate five-point dynamic profiles. We identified 81 signaling proteins, including virtually all known epidermal growth factor receptor substrates, 31 novel effectors and the time course of their activation upon epidermal growth factor stimulation. Global activation profiles provide an informative perspective on cell signaling and will be crucial to modeling signaling networks in a systems biology approach.

U2 - 10.1038/nbt1005

DO - 10.1038/nbt1005

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VL - 22

SP - 1139

EP - 1145

JO - Nature Biotechnology

JF - Nature Biotechnology

SN - 1087-0156

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