Isolation of novel Src substrates.
We have established and used a method to rapidly isolate tyrosine kinase substrates. The method entails inserting mammalian cDNA libraries into phage vectors. Protein production is induced, then plaque proteins are transferred to nitrocellulose and phosphorylated by the kinase of interest. Proof of principle for this technique was established by the isolation of a number of known Src substrates. We also implicated other known proteins as substrates for Src by this approach, and isolated a number of novel genes. Several of these are indeed Src substrates in mammalian cells. We have characterized further one of these novel substrates, Fish, which is a multi-domain adaptor protein.
Courtneidge
S A
SA
Van Andel Research Institute, 333 Bostwick, Grand Rapids, MI 49503, USA. Sara.Courtneidge@vai.org
D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000502:physiology*; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000502:physiology*; D018076:DNA, Complementary / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D008954:Models, Biological; D010766:Phosphorylation; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D015398:Signal Transduction; D019061:src-Family Kinases / Q000737:chemistry* / Q000378:metabolism*
D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D018076:DNA, Complementary; C478582:SH3PXD2A protein, human; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases