Dissection of the insulin signaling pathway via quantitative phosphoproteomics

Marcus Krüger, Irina Kratchmarova, Blagoy Blagoev, Yu-Hua Tseng, C Ronald Kahn, Matthias Mann

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The insulin signaling pathway is of pivotal importance in metabolic diseases, such as diabetes, and in cellular processes, such as aging. Insulin activates a tyrosine phosphorylation cascade that branches to create a complex network affecting multiple biological processes. To understand the full spectrum of the tyrosine phosphorylation cascade, we have defined the tyrosine-phosphoproteome of the insulin signaling pathway, using high resolution mass spectrometry in combination with phosphotyrosine immunoprecipitation and stable isotope labeling by amino acids in cell culture (SILAC) in differentiated brown adipocytes. Of 40 identified insulin-induced effectors, 7 have not previously been described in insulin signaling, including SDR, PKCdelta binding protein, LRP-6, and PISP/PDZK11, a potential calcium ATPase binding protein. A proteomic interaction screen with PISP/PDZK11 identified the calcium transporting ATPase SERCA2, supporting a connection to calcium signaling. The combination of quantitative phosphoproteomics with cell culture models provides a powerful strategy to dissect the insulin signaling pathways in intact cells.
Original languageEnglish
JournalProceedings of the National Academy of Science of the United States of America
Volume105
Issue number7
Pages (from-to)2451-2456
Number of pages5
ISSN0027-8424
DOIs
Publication statusPublished - 2008

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Insulin
Tyrosine
Brown Adipocytes
Isotope Labeling
Phosphotyrosine
Metabolic Diseases
Amino Acids

Keywords

  • Adipocytes, Brown
  • Amino Acids
  • Animals
  • Biological Transport
  • Calcium
  • Carrier Proteins
  • Cell Differentiation
  • Cells, Cultured
  • Insulin
  • Mice
  • Myosins
  • Phosphorylation
  • Protein Binding
  • Proteomics
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Signal Transduction

Cite this

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title = "Dissection of the insulin signaling pathway via quantitative phosphoproteomics",
abstract = "The insulin signaling pathway is of pivotal importance in metabolic diseases, such as diabetes, and in cellular processes, such as aging. Insulin activates a tyrosine phosphorylation cascade that branches to create a complex network affecting multiple biological processes. To understand the full spectrum of the tyrosine phosphorylation cascade, we have defined the tyrosine-phosphoproteome of the insulin signaling pathway, using high resolution mass spectrometry in combination with phosphotyrosine immunoprecipitation and stable isotope labeling by amino acids in cell culture (SILAC) in differentiated brown adipocytes. Of 40 identified insulin-induced effectors, 7 have not previously been described in insulin signaling, including SDR, PKCdelta binding protein, LRP-6, and PISP/PDZK11, a potential calcium ATPase binding protein. A proteomic interaction screen with PISP/PDZK11 identified the calcium transporting ATPase SERCA2, supporting a connection to calcium signaling. The combination of quantitative phosphoproteomics with cell culture models provides a powerful strategy to dissect the insulin signaling pathways in intact cells.",
keywords = "Adipocytes, Brown, Amino Acids, Animals, Biological Transport, Calcium, Carrier Proteins, Cell Differentiation, Cells, Cultured, Insulin, Mice, Myosins, Phosphorylation, Protein Binding, Proteomics, Sarcoplasmic Reticulum Calcium-Transporting ATPases, Signal Transduction",
author = "Marcus Kr{\"u}ger and Irina Kratchmarova and Blagoy Blagoev and Yu-Hua Tseng and Kahn, {C Ronald} and Matthias Mann",
year = "2008",
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language = "English",
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journal = "Proceedings of the National Academy of Sciences of the United States of America",
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}

Dissection of the insulin signaling pathway via quantitative phosphoproteomics. / Krüger, Marcus; Kratchmarova, Irina; Blagoev, Blagoy; Tseng, Yu-Hua; Kahn, C Ronald; Mann, Matthias.

In: Proceedings of the National Academy of Science of the United States of America, Vol. 105, No. 7, 2008, p. 2451-2456.

Research output: Contribution to journalJournal articleResearchpeer-review

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T1 - Dissection of the insulin signaling pathway via quantitative phosphoproteomics

AU - Krüger, Marcus

AU - Kratchmarova, Irina

AU - Blagoev, Blagoy

AU - Tseng, Yu-Hua

AU - Kahn, C Ronald

AU - Mann, Matthias

PY - 2008

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N2 - The insulin signaling pathway is of pivotal importance in metabolic diseases, such as diabetes, and in cellular processes, such as aging. Insulin activates a tyrosine phosphorylation cascade that branches to create a complex network affecting multiple biological processes. To understand the full spectrum of the tyrosine phosphorylation cascade, we have defined the tyrosine-phosphoproteome of the insulin signaling pathway, using high resolution mass spectrometry in combination with phosphotyrosine immunoprecipitation and stable isotope labeling by amino acids in cell culture (SILAC) in differentiated brown adipocytes. Of 40 identified insulin-induced effectors, 7 have not previously been described in insulin signaling, including SDR, PKCdelta binding protein, LRP-6, and PISP/PDZK11, a potential calcium ATPase binding protein. A proteomic interaction screen with PISP/PDZK11 identified the calcium transporting ATPase SERCA2, supporting a connection to calcium signaling. The combination of quantitative phosphoproteomics with cell culture models provides a powerful strategy to dissect the insulin signaling pathways in intact cells.

AB - The insulin signaling pathway is of pivotal importance in metabolic diseases, such as diabetes, and in cellular processes, such as aging. Insulin activates a tyrosine phosphorylation cascade that branches to create a complex network affecting multiple biological processes. To understand the full spectrum of the tyrosine phosphorylation cascade, we have defined the tyrosine-phosphoproteome of the insulin signaling pathway, using high resolution mass spectrometry in combination with phosphotyrosine immunoprecipitation and stable isotope labeling by amino acids in cell culture (SILAC) in differentiated brown adipocytes. Of 40 identified insulin-induced effectors, 7 have not previously been described in insulin signaling, including SDR, PKCdelta binding protein, LRP-6, and PISP/PDZK11, a potential calcium ATPase binding protein. A proteomic interaction screen with PISP/PDZK11 identified the calcium transporting ATPase SERCA2, supporting a connection to calcium signaling. The combination of quantitative phosphoproteomics with cell culture models provides a powerful strategy to dissect the insulin signaling pathways in intact cells.

KW - Adipocytes, Brown

KW - Amino Acids

KW - Animals

KW - Biological Transport

KW - Calcium

KW - Carrier Proteins

KW - Cell Differentiation

KW - Cells, Cultured

KW - Insulin

KW - Mice

KW - Myosins

KW - Phosphorylation

KW - Protein Binding

KW - Proteomics

KW - Sarcoplasmic Reticulum Calcium-Transporting ATPases

KW - Signal Transduction

U2 - 10.1073/pnas.0711713105

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

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