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PMID: 10924144 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Attractant regulation of the aspartate receptor-kinase complex: limited cooperative interactions between receptors and effects of the receptor modification state.

Biochemistry ·Vol. 39 ·No. 31 ·2000-08-08 ·Pages 9486-93

Bornhorst JA, Falke JJ

Abstract

The manner by which the bacterial chemotaxis system responds to a wide range of attractant concentrations remains incompletely understood. In principle, positive cooperativity between chemotaxis receptors could explain the ability of bacteria to respond to extremely low attractant concentrations. By utilizing an in vitro receptor-coupled kinase assay, the attractant-dependent response curve has been measured for the Salmonella typhimurium aspartate chemoreceptor. The attractant chosen, alpha-methyl aspartate, was originally used to quantitate high receptor sensitivity at low attractant concentrations by Segall, Block, and Berg [(1986) Proc. Natl. Acad. Sci. U.S.A. 83, 8987-8991]. The attractant response curve exhibits limited positive cooperativity, yielding a Hill coefficient of 1.7-2.4, and this Hill coefficient is relatively independent of both the receptor modification state and the mole ratio of CheA to receptor. These results disfavor models in which there are strong cooperative interactions between large numbers of receptor dimers in an extensive receptor array. Instead, the results are consistent with cooperative interactions between a small number of coupled receptor dimers. Because the in vitro receptor-coupled kinase assay utilizes higher than native receptor densities arising from overexpression, the observed positive cooperativity may overestimate that present in native receptor populations. Such positive cooperativity between dimers is fully compatible with the negative cooperativity previously observed between the two symmetric ligand binding sites within a single dimer. The attractant affinity of the aspartate receptor is found to depend on the modification state of its covalent adaptation sites. Increasing the the level of modification decreases the apparent attractant affinity at least 10-fold in the in vitro receptor-coupled kinase assay. This observation helps explain the ability of the chemotaxis pathway to respond to a broad range of attractant concentrations in vivo.

MeSH Terms
Adaptation, Physiological Aspartate Kinase/metabolism,physiology Bacterial Proteins/genetics,metabolism Chemotaxis/physiology Escherichia coli/genetics,metabolism Escherichia coli Proteins Histidine Kinase Membrane Proteins/genetics,metabolism Methyl-Accepting Chemotaxis Proteins Models, Biological Models, Chemical Mutagenesis, Site-Directed Protein Kinases/metabolism,physiology Receptors, Amino Acid/metabolism,physiology Salmonella typhimurium/enzymology,genetics,metabolism,physiology Signal Transduction
Chemicals
Bacterial Proteins CheW protein, E coli Escherichia coli Proteins Membrane Proteins Methyl-Accepting Chemotaxis Proteins Receptors, Amino Acid aspartic acid receptor CheW protein, Bacteria Protein Kinases Histidine Kinase cheA protein, E coli Aspartate Kinase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bornhorst J A
Department of Chemistry and Biochemistry, University of Colorado at Boulder, 80309-0215, USA.
Falke J J
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Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
0006-2960
DOI
10.1021/bi0002737
Published
2000-08-08
Pages
9486-93
Language
English
Region
United States
NLM ID
0370623
PMCID
PMC2890267
Subset
IM
Grants
NIGMS NIH HHS · R01 GM040731 · United States
NIGMS NIH HHS · R01 GM040731-13 · United States
NIGMS NIH HHS · GM R01-40731 · United States
External Links
PubMed source DOI: 10.1021/bi0002737 PMC Full Text View journal details
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