In vitro methylation of bacterial chemotaxis proteins: characterization of protein methyltransferase activity in crude extracts of Salmonella typhimurium.
A specific in vitro assay was developed for the protein carboxyl methyltransferase that is involved in the chemotactic behavior of Salmonella typhimurium. This cytosolic enzyme catalyzes an S-adenosyl-L-methionine-dependent methyl esterification of glutamyl residues on a class of 60,000-dalton inner-membrane proteins. The activity was found to display a pH optimum of 6.5 and be sensitive to the concentration of salts in the assay medium. No detectable activity was found towards a variety of other proteins which serve as substrates for mammalian and other bacterial carboxyl methyltransferases. This assay was used to quantitate the methylation of the 60,000-dalton methyl-accepting proteins in response to chemoeffectors. Small but reproducible concentration-dependent changes in the initial rates of in vitro methylation were observed with chemotactic attractants and repellents. The specific methyltransferase activity was found to be absent in several mutants in flagellar synthesis (fla-), suggesting that the synthesis of this enzyme is coordinately regulated with that of flagellin and basal bodies. The hydrodynamic properties of the enzyme in crude extracts were determined by gel filtration and sucrose velocity gradient centrifugation, and a native molecular weight of 41,000 was calculated from these data.
D001426:Bacterial Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000032:analysis; D002630:Chemotactic Factors / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; D008745:Methylation; D008970:Molecular Weight; D009994:Osmolar Concentration; D011496:Protein Methyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000201:enzymology*; D013045:Species Specificity
D001426:Bacterial Proteins; D002630:Chemotactic Factors; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; D011496:Protein Methyltransferases