J. Biol. Chem., Vol. 259, Issue 19, 11828-11835, 10, 1984

Stimulus-induced changes in methylesterase activity during chemotaxis in Escherichia coli

MR Kehry, TG Doak and FW Dahlquist

Responses of Escherichia coli to chemical stimuli are mediated by a family of sensory transducer proteins. These transmembrane proteins detect stimuli and convey sensory information to the flagella. Behavioral adaptation to environmental changes is correlated with the reversible methylation of the transducer proteins on several (4 to 6) specific glutamic acid residues to form methyl esters. We have assayed the activity of the chemotaxis-specific methylesterase, the product of the cheB gene, by measuring the product of the demethylation reaction, methanol, using a modification of a previous method (Toews, M.L., Goy, M.F., Springer, M.S., and Adler, J. (1979) Proc. Natl. Acad. Sci. U.S.A. 76, 5544-5548; Terwilliger, T.C., Bogonez, E., Wang, E.A., and Koshland, D. E., Jr. (1983) J. Biol. Chem. 258, 9608-9611). In this study, we establish the use of a sensitive flow apparatus for measuring stimulus-induced changes in methylesterase activity of intact E. coli cells. Responses to positive and negative stimuli consist of transient decrease and increases in methylesterase activity, respectively. In addition, chase experiments demonstrated that all assayable methyl groups are nearly equivalent. The results are not consistent with the view that the methyl groups put on the transducer proteins as a result of a positive stimulus are the first to be removed when that stimulus is withdrawn. It seems more likely that recently added methyl groups become part of a pool of kinetically equivalent members, any of which can be removed when the stimulus is withdrawn. The flow measurements provide a powerful and simple biochemical assay that complements behavioral studies of chemotaxis.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				B. B. Stephens, S. N. Loar, and G. Alexandre Role of CheB and CheR in the Complex Chemotactic and Aerotactic Pathway of Azospirillum brasilense J. Bacteriol., July 1, 2006; 188(13): 4759 - 4768. [Abstract] [Full Text] [PDF]

				F. M. Antommattei, J. B. Munzner, and R. M. Weis Ligand-Specific Activation of Escherichia coli Chemoreceptor Transmethylation J. Bacteriol., November 15, 2004; 186(22): 7556 - 7563. [Abstract] [Full Text] [PDF]

				H. Szurmant and G. W. Ordal Diversity in Chemotaxis Mechanisms among the Bacteria and Archaea Microbiol. Mol. Biol. Rev., June 1, 2004; 68(2): 301 - 319. [Abstract] [Full Text] [PDF]

				A. N. Barnakov, L. A. Barnakova, and G. L. Hazelbauer Allosteric Enhancement of Adaptational Demethylation by a Carboxyl-terminal Sequence on Chemoreceptors J. Biol. Chem., October 25, 2002; 277(44): 42151 - 42156. [Abstract] [Full Text] [PDF]

				A. C. Martin, G. H. Wadhams, D. S. H. Shah, S. L. Porter, J. C. Mantotta, T. J. Craig, P. H. Verdult, H. Jones, and J. P. Armitage CheR- and CheB-Dependent Chemosensory Adaptation System of Rhodobacter sphaeroides J. Bacteriol., December 15, 2001; 183(24): 7135 - 7144. [Abstract] [Full Text] [PDF]

				Z.-Y. Jiang and C. E. Bauer Component of the Rhodospirillum centenum Photosensory Apparatus with Structural and Functional Similarity to Methyl-Accepting Chemotaxis Protein Chemoreceptors J. Bacteriol., January 1, 2001; 183(1): 171 - 177. [Abstract] [Full Text]

				R. Kort, W. Crielaard, J. L. Spudich, and K. J. Hellingwerf Color-Sensitive Motility and Methanol Release Responses in Rhodobacter sphaeroides J. Bacteriol., June 1, 2000; 182(11): 3017 - 3021. [Abstract] [Full Text]

				A. N. Barnakov, L. A. Barnakova, and G. L. Hazelbauer Efficient adaptational demethylation of chemoreceptors requires the same enzyme-docking site as efficient methylation PNAS, September 14, 1999; 96(19): 10667 - 10672. [Abstract] [Full Text] [PDF]

				J. R. Kirby, M. M. Saulmon, C. J. Kristich, and G. W. Ordal CheY-dependent Methylation of the Asparagine Receptor, McpB, during Chemotaxis in Bacillus subtilis J. Biol. Chem., April 16, 1999; 274(16): 11092 - 11100. [Abstract] [Full Text] [PDF]

				M. A. Zimmer, J. Tiu, M. A. Collins, and G. W. Ordal Selective Methylation Changes on the Bacillus subtilis Chemotaxis Receptor McpB Promote Adaptation J. Biol. Chem., August 4, 2000; 275(32): 24264 - 24272. [Abstract] [Full Text] [PDF]