PMID:21858888
| Citation |
Bartelli, NL and Hazelbauer, GL (2011) Direct evidence that the carboxyl-terminal sequence of a bacterial chemoreceptor is an unstructured linker and enzyme tether. Protein Sci. 20:1856-66 |
|---|---|
| Abstract |
Sensory adaptation in bacterial chemotaxis involves reversible methylation of specific glutamyl residues on chemoreceptors. The reactions are catalyzed by a dedicated methyltransferase and dedicated methylesterase. In Escherichia coli and related organisms, control of these enzymes includes an evolutionarily recent addition of interaction with a pentapeptide activator located at the carboxyl terminus of the receptor polypeptide chain. Effective enzyme activation requires not only the pentapeptide but also a segment of the receptor polypeptide chain between that sequence and the coiled-coil body of the chemoreceptor. This segment has features consistent with a role as a flexible and presumably unstructured linker and enzyme tether, but there has been no direct information about its structure. We used site-directed spin labeling and electron paramagnetic resonance spectroscopy to characterize structural features of the carboxyl-terminal 40 residues of E. coli chemoreceptor Tar. Beginning ∼ 35 residues from the carboxyl terminus and continuing to the end of the protein, spectra of spin-labeled Tar embedded in native membranes or in reconstituted proteoliposomes, exhibited mobilities characteristic of unstructured, disordered segments. Binding of methyltransferase substantially reduced mobility for positions in or near the pentapeptide but mobility for the linker sequence remained high, being only modestly reduced in a gradient of decreasing effects for 10-15 residues, a pattern consistent with the linker providing a flexible arm that would allow enzyme diffusion within defined limits. Thus, our data identify that the carboxyl-terminal linker between the receptor body and the pentapeptide is an unstructured, disordered segment that can serve as a flexible arm and enzyme tether. |
| Links |
PubMed PMC3267950 Online version:10.1002/pro.719 |
| Keywords |
Amino Acid Sequence; Cell Membrane; Chemotaxis; Electron Spin Resonance Spectroscopy; Escherichia coli/chemistry; Escherichia coli/enzymology; Escherichia coli/physiology; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/metabolism; Methyltransferases/metabolism; Molecular Sequence Data; Protein Conformation; Proteolipids/chemistry; Receptors, Cell Surface/chemistry; Receptors, Cell Surface/metabolism; Sequence Analysis, Protein; Spin Labels |
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