Peroxovanadate Induces Tyrosine Phosphorylation of Multiple Signaling Proteins in Mouse Liver and Kidney

doi:10.1074/jbc.272.2.1263

Peroxovanadate Induces Tyrosine Phosphorylation of Multiple Signaling Proteins in Mouse Liver and Kidney*

From the Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146

^‡ To whom correspondence should be addressed:
Dept. of Biochemistry, Vanderbilt University School of Medicine, 607 Light Hall, Nashville, TN 37232-0146.
Tel.: 615-322-3318; Fax: 615-322-4349.

Abstract

The intraperitoneal injection of a vanadate/H₂O₂ mixture (peroxovanadate) into mice resulted within minutes in the appearance of numerous tyrosine-phosphorylated proteins in the liver and kidney. These effects are presumably due to the inhibition of phosphotyrosine phosphatase activity. Three of the tyrosine-phosphorylated proteins have been identified as the receptors for epidermal growth factor, insulin, and hepatocyte growth factor. The injection of peroxovanadate also enhanced the tyrosine phosphorylation of many of the proteins known to function downstream of these receptors, including SHC, signal transducer and activator of transcription (Stat) 1α,β, Stat 3, Stat 5, phospholipase C-γ, insulin receptor substrate 1, GTPase-activating protein, β-catenin, γ-catenin, p120^cas, SHP-1, and SHP-2. The administration of peroxovanadate also induced nuclear translocation of a number of tyrosine-phosphorylated Stat proteins. In addition, the global effects on tyrosine phosphorylation permitted the detection of a number of novel intracellular protein interactions, including an association of Tyk2 with β-catenin. The in situ administration of peroxovanadate may prove useful in the search for novel tyrosine-phosphorylated proteins and the identification of new interactions between previously identified tyrosine-phosphorylated substrates.

Footnotes

↵* This work was supported by United States Public Health Service Grant HD-00700. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

IRS-1

insulin receptor substrates

EGF

epidermal growth factor

PAGE

polyacrylamide gel electrophoresis

RC20H

horseradish peroxidase-conjugated recombinant antibody fragment specific for phosphotyrosine

PBS

Dulbecco's calcium-free magnesium-free phosphate-buffered saline

Stat

signal transducer and activator of transcription

PLC-γ

phospholipase C-γ.
- Received August 31, 1996.
- Revision received November 4, 1996.