Location of the receptor-interaction site on CheB, the methylesterase response regulator of bacterial chemotaxis.
Sensory adaptation in bacterial chemotaxis is mediated by covalent modification of chemoreceptors, specifically methylation and demethylation of glutamates catalyzed by methyltransferase CheR and methylesterase CheB. The methylesterase is a two-domain response regulator in which phosphorylation of the regulatory domain enhances activity of the catalytic domain. In Escherichia coli and Salmonella typhimurium, a crucial determinant of efficient methylation and demethylation is a specific pentapeptide sequence at the chemoreceptor carboxyl terminus, a position distant from sites of enzymatic action. Each enzyme binds pentapeptide, but the site of binding has been located only for CheR. Here we locate the pentapeptide-binding site on CheB by assessing catalytic activity and pentapeptide binding of CheB fragments, protection of CheB from proteolysis by pentapeptide, and interference with pentapeptide-CheB interaction by a CheB segment. The results place the binding site near the hinge between regulatory and catalytic domains, in a segment spanning the carboxyl-terminal end of the regulatory domain and the beginning of the linker that stretches to the catalytic domain. This location is quite different from the catalytic domain location of the pentapeptide-binding site on CheR and is likely to reflect the rather different ways in which pentapeptide binding enhances enzymatic action for the methyltransferase and the methylesterase.
Barnakov
A N
AN
Department of Biochemistry, University of Missouri-Columbia, Columbia, Missouri 65211, USA.
Barnakova
L A
LA
Hazelbauer
G L
GL
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D020134:Catalytic Domain; D002630:Chemotactic Factors / Q000737:chemistry / Q000378:metabolism; D002633:Chemotaxis / Q000502:physiology*; D004926:Escherichia coli / Q000502:physiology*; D018698:Glutamic Acid / Q000378:metabolism; D008780:Methyltransferases / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010449:Peptide Mapping; D017433:Protein Structure, Secondary; D012486:Salmonella typhimurium / Q000502:physiology*; D014357:Trypsin
D001426:Bacterial Proteins; D002630:Chemotactic Factors; D009842:Oligopeptides; D010446:Peptide Fragments; C069656:CheB protein, Bacteria; D018698:Glutamic Acid; D008780:Methyltransferases; C038706:chemotaxis methyltransferase; D014357:Trypsin