A RECEPTOR TYROSINE KINASE TARGET PROTEIN cDNA CLONING METHOD AND hGRB PROTEINS

摘要

A novel expression cloning method for detection and identification of proteins capable of binding to tyrosine-phosphorylated domains of receptor tyrosine kinases is based on the use of a novel probe. This probe comprises an amino acid sequence derived from the tyrosine-phosphorylated portion of the receptor molecule, or a functional derivative thereof, and has at least one phosphorylated tyrosine residue, lacks the tyrosine kinase portion of the receptor, and is detectably labeled. Also disclosed are a method for preparing the probe, a method for mapping to a chromosome a gene encoding a protein capable of binding to tyrosine-phosphorylated domains of receptor tyrosine kinases, and a method for purifying such a protein with the probe. Two proteins discovered using the above cloning method, GRB-1 and GRB-2, DNA encoding these proteins, and methods for detecting these proteins are also disclosed.