Peroxynitrite induces covalent dimerization of epidermal growth factor receptors in A431 epidermoid carcinoma cells

Albert Van Der Vliet, Milena Hristova, Carroll E Cross, Jason P. Eiserich, Tzipora Goldkorn

Research output: Contribution to journalArticle

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Abstract

Irreversible tyrosine modifications by inflammatory oxidants such as peroxynitrite (ONOO-) can affect signal transduction pathways involving tyrosine phosphorylation. The epidermal growth factor receptor (EGFR), a member of the c-ErbB receptor tyrosine kinase family, is involved in regulation of epithelial cell growth and differentiation, and possible modulation of EGFR-dependent signaling by ONOO- was studied. Exposure of epidermoid carcinoma A431 cells to 0.1-1.0 mM ONOO- resulted in tyrosine nitration on EGFR and other proteins but did not significantly affect EGFR tyrosine autophosphorylation. A high molecular mass tyrosine-phosphorylated protein (~340 kDa) was detected in A431 cell lysates after exposure to ONOO-, most likely representing a covalently dimerized form of EGFR, based on immunoprecipitation and/or immunoblotting with α-EGFR antibodies and co- migration with ligand-induced EGFR dimers cross-linked with 1-ethyl-3-(3- dimethylaminopropyl)carbodiimide. Covalent EGFR dimerization by ONOO- probably involved intermolecular dityrosine cross-linking and was enhanced after receptor activation with epidermal growth factor. Furthermore, irreversibly cross-linked EGFR was more extensively tyrosine-phosphorylated compared with the monomeric form, indicating that ONOO- preferentially cross-links activated EGFR. Exposure of A431 cells to ONOO- markedly reduced the kinetics of tyrosine phosphorylation of a downstream EGFR substrate, phospholipase C-γ1, which may be related to covalent alterations in EGFR. Alteration of EGFR signaling by covalent EGFR dimerization by inflammatory oxidants such as ONOO- may affect conditions of increased EGFR activation such as epithelial repair or tumorigenesis.

Original languageEnglish (US)
Pages (from-to)31860-31866
Number of pages7
JournalJournal of Biological Chemistry
Volume273
Issue number48
DOIs
StatePublished - Nov 27 1999

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Peroxynitrous Acid
Dimerization
Epidermal Growth Factor Receptor
Squamous Cell Carcinoma
Cells
Tyrosine
Phosphorylation
ErbB Receptors
Oxidants
Ethyldimethylaminopropyl Carbodiimide
Chemical activation
Nitration
Signal transduction
Cell growth
Molecular mass
Type C Phospholipases
Immunoprecipitation
Immunoblotting
Epidermal Growth Factor
Dimers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Peroxynitrite induces covalent dimerization of epidermal growth factor receptors in A431 epidermoid carcinoma cells. / Van Der Vliet, Albert; Hristova, Milena; Cross, Carroll E; Eiserich, Jason P.; Goldkorn, Tzipora.

In: Journal of Biological Chemistry, Vol. 273, No. 48, 27.11.1999, p. 31860-31866.

Research output: Contribution to journalArticle

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