The role of individual SH2 domains in mediating association of phospholipase C-γ1 with the activated EGF receptor

Ansuman Chattopadhyay, Manuela Vecchi, Qun Sheng Ji, Raymond Mernaugh, Graham Carpenter

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

The two SH2 (Src homology domain 2) domains present in phospholipase C- γ1 (PLC-γ1) were assayed for their capacities to recognize the five autophosphorylation sites in the epidermal growth factor receptor. Plasmon resonance and immunological techniques were employed to measure interactions between SH2 fusion proteins and phosphotyrosine-containing peptides. The N- SH2 domain recognized peptides in the order of pY1173 > pY992 > pY1068 > pY1148 >> pY1086, while the C-SH2 domain recognized peptides in the order of pY992 > pY1068 > pY1148 >> pY1086 and pY1173. The major autophosphorylation site, pY1173, was recognized only by the N-SH2 domain. Contributions of the N-SH2 and C-SH2 domains to the association of the intact PLC-γ1 molecule with the activated epidermal growth factor (EGF) receptor were assessed in vivo. Loss of function mutants of each SH2 domain were produced in a full- length epitope-tagged PLC-γ1. After expression of the mutants, cells were treated with EGF and association of exogenous PLC-γ1 with EGF receptors was measured. In this context the N-SH2 is the primary contributor to PLC-γ1 association with the EGF receptor. The combined results suggest an association mechanism involving the N-SH2 domain and the pY1173 autophosphorylation site as a primary event and the C-SH2 domain and the pY992 autophosphorylation site as a secondary event.

Original languageEnglish
Pages (from-to)26091-26097
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number37
DOIs
Publication statusPublished - Sep 10 1999

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src Homology Domains
Type C Phospholipases
Epidermal Growth Factor Receptor
Peptides
Phosphotyrosine
Epidermal Growth Factor
Epitopes
Fusion reactions
Immunologic Techniques
Molecules
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

The role of individual SH2 domains in mediating association of phospholipase C-γ1 with the activated EGF receptor. / Chattopadhyay, Ansuman; Vecchi, Manuela; Ji, Qun Sheng; Mernaugh, Raymond; Carpenter, Graham.

In: Journal of Biological Chemistry, Vol. 274, No. 37, 10.09.1999, p. 26091-26097.

Research output: Contribution to journalArticle

Chattopadhyay, Ansuman ; Vecchi, Manuela ; Ji, Qun Sheng ; Mernaugh, Raymond ; Carpenter, Graham. / The role of individual SH2 domains in mediating association of phospholipase C-γ1 with the activated EGF receptor. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 37. pp. 26091-26097.
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