Identification of signalling components in tyrosine kinase cascades using phosphopeptide affinity chromatography

Biochemical and Biophysical Research Communications
G DuménilS Pellegrini

Abstract

Various methods are now available to identify the molecular partners of the component of a signal transduction pathway. Some interactions, however, can be technically difficult to detect because they depend upon transient tyrosine phosphorylation. Here, we present a simple affinity chromatography approach based on synthetic phosphopeptides to purify potential partners of phosphotyrosine-containing proteins. With this approach, we confirm the previously characterized interaction between Grb2 and the EGF receptor, and we identify novel partners of the IGF-1 receptor and of the JAK proteins. Methenyltetrahydrofolate synthetase (MTHFS) was identified as a potential mediator of IGF-1R dependent transformation. P85alpha, the regulatory subunit of PI3 kinase, was identified as one of four proteins recruited by a phosphopeptide mimicking a motif conserved in all JAK family members.

References

Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Dec 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·R R RousselA P Laudano
Apr 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·W R Pearson, D J Lipman
Nov 17, 1995·The Journal of Biological Chemistry·G WolfS E Shoelson
Aug 1, 1994·Molecular and Cellular Biology·A G BatzerJ Schlessinger
Dec 16, 1994·Science·W M Kavanaugh, L T Williams
Feb 16, 1995·Nature·T Pawson
Jan 1, 1995·Annual Review of Biochemistry·C Schindler, J E Darnell
Mar 12, 1993·Cell·Z SongyangR J Lechleider
Feb 1, 1995·Trends in Genetics : TIG·J N Ihle, I M Kerr
Feb 1, 1994·Nature Genetics·S F AltschulJ C Wootton
Jan 18, 1994·Proceedings of the National Academy of Sciences of the United States of America·D W RoseJ M Olefsky
Sep 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·O SilvennoinenJ N Ihle
Jul 16, 1996·Biochemical and Biophysical Research Communications·D K Fuhrer, Y C Yang

❮ Previous
Next ❯

Related Concepts

Related Feeds

AKT Pathway

This feed focuses on the AKT serine/threonine kinase, which is an important signaling pathway involved in processes such as glucose metabolism and cell survival.