Kinetics of receptor modification. The multiply methylated aspartate receptors involved in bacterial chemotaxis.
A method for determining the extent of methyl esterification of each of the four potential sites on the aspartate receptors involved in chemotaxis in Escherichia coli and Salmonella typhimurium is presented. In this procedure, radioactive methyl esters are incorporated into the receptors, the receptors are cleaved by trypsin and the V8 protease from Staphylococcus aureus, and the four fragments containing sites of methylation are separated by high performance liquid chromatography. Using this technique, we find that the rate of methyl esterification increases at all four sites after stimulation with the "attractant" aspartate, suggesting that all four sites of modification are involved in adaptation to aspartate. We also find that the rate of methyl esterification at each site is correlated with the homology between the protein sequence at that site and the "consensus" sequence, Glu-Glu-X-X-Ala-Thr/Ser.
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000378:metabolism*; D002633:Chemotaxis; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008745:Methylation; D010446:Peptide Fragments / Q000032:analysis; D010957:Plasmids; D017459:Receptors, Amino Acid; D017981:Receptors, Neurotransmitter / Q000302:isolation & purification / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000378:metabolism*
D010446:Peptide Fragments; D017459:Receptors, Amino Acid; D017981:Receptors, Neurotransmitter; C031959:aspartic acid receptor; D001224:Aspartic Acid