Changing reactivity of receptor carboxyl groups during bacterial sensing.

A microdistillation procedure has been developed to analyze carboxylmethylation of the Mr = 60,000 chemoreceptor proteins involved in chemotaxis of Salmonella typhimurium and Escherichia coli. Methylation levels obtained by this method are substantially higher than those reported in the literature. In highly motile strains under optimal conditions there are approximately 100,000 methylated receptor residues per cell which are entirely composed of gamma-methylglutamyl esters. Whereas with previously used methods only groups which turn over could be detected, the microdistillation assay provides absolute values. Under steady state conditions, approximately one-half the total number of methyl ester residues are continuously hydrolyzed and resynthesized, while the remainder are sequestered. A mechanism has been devised to explain the observed patterns of methyl ester synthesis and hydrolysis. According to this hypothesis, substrate glutamyl residues on the receptor are located in a restricted region near the active sites of transferase and esterase which are bound to the receptor protein. Small, stimuli-induced changes in receptor conformation effect perturbations in receptor methylation by shifting the focus of activity of the modifying enzymes from one pair of closely spaced groups to another.