title: A new type of protein methylation activated by tyrphostin A25 and vanadate.
abstract: It has been reported that S-adenosylmethionine-dependent protein methylation in rat kidney extracts can be greatly stimulated by tyrphostin A25, a tyrosine kinase inhibitor. We have investigated the nature of this stimulation. We find that addition of tyrphostin A25, in combination with the protein phosphatase inhibitor vanadate, leads to the stimulation of methylation of polypeptides of 64, 42, 40, 36, 31, and 15 kDa in cytosolic extracts of mouse kidney. The effect of tyrphostin appears to be relatively specific for the A25 species. The enhanced methylation does not represent the activity of the families of protein histidine, lysine or arginine methyltransferases, nor that of the l-isoaspartyl/d-aspartyl methyltransferase, enzymes responsible for the bulk of protein methylation in most cell types. Chemical and enzymatic analyses of the methylated polypeptides suggest that the methyl group is in an ester linkage to the protein. In heart extracts, we find a similar situation but here the stimulation of methylation is not dependent upon vanadate and an additional 18 kDa methylated species is found. In contrast, little or no stimulation of methylation is found in brain or testis extracts. This work provides evidence for a novel type of protein carboxyl methylation reaction that may play a role in signaling reactions in certain mammalian tissues.
authors.affiliation: Department of Chemistry and Biochemistry, Molecular Biology Institute, UCLA, Los Angeles, CA 90095-1569, USA.
authors.forename: Tina Branscombe
authors.initials: TB
authors.lastname: Miranda
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Lowenson
authors.forename: Steven
authors.initials: S
authors.lastname: Clarke
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D008745:Methylation; D051379:Mice; D011506:Proteins / Q000378:metabolism*; D020032:Tyrphostins / Q000494:pharmacology*; D014638:Vanadates / Q000494:pharmacology*
chemical_list: D011506:Proteins; D020032:Tyrphostins; C079836:tyrphostin 25; D014638:Vanadates