Recombinant DNA encoding a eukaryotic tyrosine kinase target pro

Details Recombinant DNA encoding a eukaryotic tyrosine kinase target pro Recombinant DNA encoding a eukaryotic tyrosine kinase target pro

1993-12-16

1997-04-08

Ulm, John

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 697, 4352523, 4353201, 530350, 536 234, 536 235, C12N 510, C12N 1512, C12N 1562

Patent

active

056186916

ABSTRACT:
A novel expression cloning method is provided for the detection, identification and purification of target proteins capable of binding at least to a tryosine-phosphorylated domain of a eukaryotic tyrosine kinase using novel peptide probes comprising an amino acid sequence substantially corresponding to a portion of a tyrosine-phosphorylated domain of a tyrosine kinase. The probe has at least one phosphorlated tyrosine residue and may be detectably labeled. Also disclosed is a method for preparing the probe, a method for mapping to a chormosome a gene encoding a protein capable of binding to tyrosine-phosphorylated domains of tyrosine kinases, and a method for purifiying such a protein with the probe. Non-limiting examples of novel proteins discovered using the above cloning method include GRB-1, GRB-2, GRB-3, GRB-4 and GRB-7, as well as nucleic acid encoding these proteins, and methods for detecting these proteins are also provided.

REFERENCES:
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Otsu, M. et al, Characterization of Two 85 kd Proteins that Associate with Receptor Tyronsine Kinases, Middle-T/pp60c-src Complexes, and Pl3-Kinase, Cell, vol. 55, pp. 91-104, Apr. 5, 1991.
Margolis, B. et al, The Tyrosine Phosphorylated Carboxyterminus of the EGF Receptor is a Binding site for GAP and PLC-y, The EMBO Journal, vol. 9, No. 13, pp. 4375-4380, 1990.
MacGregor, P.F. et al, Direct Cloning of Leucine Zipper Proteins: Jun Binds Cooperatively to the CRE with CRE-BP1, Oncogene, vol. 5, pp. 451-458, 1990.
Ullrich, A. et al, Signal Transduction by Receptors with Tyrosine Kinase Activity, Cell, vol. 61, pp. 203-212, Apr. 20, 1990.
Skolnik, E.Y. et al, Cloning of P13 Kinase-Associated p85 Utilizing a Novel Method for Expression/Cloning of Target Proteins for Receptor Tyrosine Kinases, Cell, vol. 65, pp. 83-90, Apr. 5, 1991.

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