Nitric oxide reversibly inhibits the epidermal growth factor receptor tyrosine kinase
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Title: | Nitric oxide reversibly inhibits the epidermal growth factor receptor tyrosine kinase |
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Authors: | Estrada Cerquera, Carmen | Gómez Torres, Carmen | Martín-Nieto, José | Frutos Herranz, Trinidad de | Jiménez Martínez, Amparo | Villalobo Polo, Antonio |
Research Group/s: | Genética Humana y de Mamíferos |
Center, Department or Service: | Universidad de Alicante. Departamento de Fisiología, Genética y Microbiología | Instituto de Investigaciones Biomédicas | Universidad Autónoma de Madrid. Departamento de Fisiología |
Keywords: | Nitric oxide | Epidermal growth factor receptor | Tyrosine kinase |
Knowledge Area: | Genética | Fisiología |
Date Created: | 2-Jan-1997 |
Issue Date: | 1-Sep-1997 |
Publisher: | Portland Press |
Citation: | ESTRADA CERQUERA, Carmen, et al. "Nitric oxide reversibly inhibits the epidermal growth factor receptor tyrosine kinase". Biochemical Journal. Vol. 326, Part 2 (Sept. 1997). ISSN 0264-6021, pp. 369-376 |
Abstract: | Although it has been demonstrated that NO inhibits the proliferation of different cell types, the mechanisms of its anti-mitotic action are not well understood. In this work we have studied the possible interaction of NO with the epidermal growth factor receptor (EGFR), using transfected fibroblasts which overexpress the human EGFR. The NO donors S-nitroso-N-acetylpenicillamine (SNAP), 1,1-diethyl-2-hydroxy-2-nitrosohydrazine (DEA-NO) and N-{4-[1-(3-aminopropyl)-2-hydroxy-2-nitrosohydrazino]butyl}propane-1,3-diamine (DETA-NO) inhibited DNA synthesis of fibroblasts growing in the presence of fetal calf serum, epidermal growth factor (EGF) or EGF plus insulin, as assessed by [methyl-3H]thymidine incorporation. Neither 8-bromo-cGMP nor the cGMP-phosphodiesterase inhibitor zaprinast mimicked this effect, suggesting that NO is unlikely to inhibit cell proliferation via a cGMP-dependent pathway. SNAP, DEA-NO and DETA-NO also inhibited the transphosphorylation of the EGFR and its tyrosine kinase activity toward the exogenous substrate poly-L-(Glu-Tyr), as measured in permeabilized cells using [γ-32P]ATP as phosphate donor. In contrast, 3-[morpholinosydnonimine hydrochloride] (SIN-1), a peroxynitrite-forming compound, did not significantly inhibit either DNA synthesis or the EGFR tyrosine kinase activity. The inhibitory action of DEA-NO on the EGFR tyrosine kinase was prevented by haemoglobin, an NO scavenger, but not by superoxide dismutase, and was reversed by dithiothreitol. The binding of EGF to its receptor was unaffected by DEA-NO. The inhibitory action of DEA-NO on the EGF-dependent transphosphorylation of the receptor was also demonstrated in intact cells by immunoblot analysis using an anti-phosphotyrosine antibody. Taken together, these results suggest that NO, but not peroxynitrite, inhibits in a reversible manner the EGFR tyrosine kinase activity by S-nitrosylation of the receptor. |
Sponsor: | This work was supported by a grant to A.V. from the Comisión Interministerial de Ciencia y Tecnología (SAF96-0035) and a grant to C.E. from the Fondo de Investigaciones Sanitarias de la Seguridad Social (97/2054). J.M.-N. is the recipient of a postdoctoral research contract from the Ministerio de Educación y Ciencia. |
URI: | http://hdl.handle.net/10045/9735 |
ISSN: | 0264-6021 (Print) | 1470-8728 (Online) |
Language: | eng |
Type: | info:eu-repo/semantics/article |
Rights: | The final version of record is available at http://www.biochemj.org/ |
Peer Review: | si |
Publisher version: | http://www.biochemj.org/ |
Appears in Collections: | INV - GHM - Artículos de Revistas |
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Estrada et al. 1997 BJ.pdf | Versión final (acceso restringido) | 368,83 kB | Adobe PDF | Open Request a copy |
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