The Grb2-mSos1 complex binds phosphopeptides with higher affinity than Grb2.

The Journal of Biological Chemistry
Y M ChookT Pawson

Abstract

Epidermal growth factor (EGF) stimulation leads to autophosphorylation of the epidermal growth factor receptor (EGFR) and tyrosine phosphorylation of Shc. The Grb2 SH2 domain binds to Tyr1068 of EGFR and Tyr317 of Shc while its SH3 domains bind to mSos1. Therefore, EGF treatment potentially results in the formation of several multimeric signaling complexes, including EGFR-Grb2-mSos1, EGFR-Shc-Grb2-mSos1, and Shc-Grb2-mSos1, linking the receptor to activation of the Ras GTPase. We have purified Grb2, mSos1, and the Grb2-mSos1 complex to high homogeneity, and used these isolated proteins to obtain binding affinities of mSos1 for Grb2 and of either Grb2 or Grb2-mSos1 for phosphotyrosine-containing peptides. mSos1 bound Grb2 with a KD of 0.4 microM; the stoichiometry of the Grb2-mSos1 complex was 1:1. An EGFR-derived phosphopeptide bound Grb2 with a KD of 0.7 microM, whereas the Shc-derived phosphopeptide bound Grb2 with a KD of 0.2 microM. Since Grb2 exists in a stable complex with mSos1, and both proteins can exist in a constitutive complex in unstimulated cells, we performed phosphopeptide binding studies on the Grb2-mSos1 complex to gain a better understanding of binding events in the intact cell. Grb2-mSos1 bound to both EGFR-...Continue Reading

References

Jul 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·D BowtellP Senior
Apr 4, 1969·Analytical Biochemistry·J Babul, E Stellwagen
Aug 1, 1994·Molecular and Cellular Biology·A G BatzerJ Schlessinger
Feb 1, 1995·Molecular and Cellular Biology·K S RavichandranS J Burakoff
Mar 12, 1993·Cell·Z SongyangR J Lechleider
Apr 14, 1995·Science·S MaignanA Ducruix
Mar 1, 1994·Molecular and Cellular Biology·G J PronkJ L Bos
Feb 1, 1994·Current Opinion in Genetics & Development·F McCormick

❮ Previous
Next ❯

Citations

Feb 11, 1999·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·A A ButlerD LeRoith
Aug 15, 2006·Nature Structural & Molecular Biology·Jon C D HoutmanLawrence E Samelson
Oct 21, 2006·Systems Biology·N I MarkevichB N Kholodenko
Aug 14, 2012·Proceedings of the National Academy of Sciences of the United States of America·Dongmyung OhJi Yu
May 5, 2000·The Journals of Gerontology. Series A, Biological Sciences and Medical Sciences·D HutterY Liu
May 13, 2010·BMC Systems Biology·Ming-yu HsiehBridget S Wilson
Jul 5, 2012·The Journal of Chemical Physics·Jin Yang, John E Pearson
Nov 10, 2007·Proceedings of the National Academy of Sciences of the United States of America·Miki MorimatsuYasushi Sako
Feb 17, 2001·Journal of Molecular Biology·S YuzawaF Inagaki
Nov 8, 2011·Journal of Proteomics·Simone LemeerBernhard Kuster
Nov 15, 2012·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Rebekah R Bartelt, Jon C D Houtman
Jun 17, 2015·Biochimica Et Biophysica Acta·Rebekah R BarteltJon C D Houtman
Jul 3, 2016·Proceedings of the National Academy of Sciences of the United States of America·William Y C HuangJay T Groves
Nov 5, 1997·The Journal of Biological Chemistry·M AnafiT Pawson
Jan 17, 1997·The Journal of Biological Chemistry·T J KubiseskiT Pawson
Oct 9, 1999·The Journal of Biological Chemistry·B N KholodenkoJ B Hoek
Feb 7, 1998·The Journal of Biological Chemistry·J W ThomasM D Schaller
Mar 19, 2020·Communications Biology·Nikolaos PatsoukisVassiliki A Boussiotis
Dec 18, 2003·The Journal of Biological Chemistry·Waltraud X Schulze, Matthias Mann
May 29, 1997·Biochemical and Biophysical Research Communications·M RojasY Z Lin
Aug 26, 1998·Biochemical and Biophysical Research Communications·L J GrahamE Bonvini
Jul 8, 2021·The Biochemical Journal·Neda S Kazemein JasemiMohammad R Ahmadian
Apr 5, 2014·Biochemistry·Noga KozerAndrew H A Clayton
Jan 24, 2020·Journal of the American Chemical Society·Tsung-Jen LiaoDavid Fushman
Sep 22, 2011·Science Signaling·Susana R Neves

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.

Advanced Imaging of Cellular Signaling

Cell signaling is a vital mechanism for communication within cells and outside with the environment. Several different signaling pathways have been found and advanced imaging techniques are being developed to visualize the molecules involved in these signaling pathways. Find the latest research in advanced imaging of cellular signaling here.