Investigating the substrate specificity of the HER2/Neu tyrosine kinase using peptide libraries

Chan, P. M., Nestler, H. P., Miller, W. T. (November 2000) Investigating the substrate specificity of the HER2/Neu tyrosine kinase using peptide libraries. Cancer Letters, 160 (2). pp. 159-169. ISSN 0304-3835

URL: http://www.ncbi.nlm.nih.gov/pubmed/11053645
DOI: 10.1016/S0304-3835(00)00581-4

Abstract

The product of the HER2/Neu oncogene is a receptor tyrosine kinase that is amplified in 25-30% of human primary breast tumors. In this project? we have isolated the HER2/Neu kinase from Sf9 cells infected with a baculovirus expression vector. We probed the substrate specificity of the HER2/Neu kinase using two peptide libraries: (1) a soluble peptide library containing three degenerate positions N-terminal to tyrosine; and (2) a bead-supported combinatorial library possessing sis degenerate positions at P - 1, P - 2, P - 3, P + 1, P + 2, and P + 3. We identified four novel substrate sequences for HER2/Neu from the two peptide libraries. We synthesized these peptides as individual sequences and measured steady-state kinetic properties for phosphorylation by HER2/Neu. One of the peptides, AAEEIYAARRG, is the best synthetic peptide substrate reported to date for HER2/Neu. All of the sequences bear a resemblance to sites of autophosphorylation on HER2/Neu and related epidermal growth factor (EGF) receptor family tyrosine kinases.

Item Type: Paper
Uncontrolled Keywords: HER2/Neu tyrosine kinase peptide library substrate specificity RETINOBLASTOMA GENE-PRODUCT HUMAN-BREAST-CANCER NEU-ONCOGENE AUTOPHOSPHORYLATION SITES PROTEIN-KINASES EGF RECEPTOR IDENTIFICATION GROWTH TRANSFORMATION MOTIFS
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > HER2
diseases & disorders > cancer > cancer types > breast cancer
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase > tyrosine kinase
CSHL Authors:
Communities: CSHL labs
Depositing User: Matt Covey
Date: November 2000
Date Deposited: 30 Jan 2014 17:01
Last Modified: 30 Jan 2014 17:01
Related URLs:
URI: http://repository.cshl.edu/id/eprint/29408

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