title: Modular structure of the Rhizobium meliloti DctB protein.
abstract: To investigate the modular structure of the Rhizobium meliloti dicarboxylic acid sensor protein, DctB, three truncated DctB proteins (DctB4, DctB5 and DctB4G) were constructed, overproduced in Escherichia coli and purified. The DctB4G protein was composed of 446 amino acids of the DctB C-terminus and displayed strong autophosphorylation activity in vitro. This activity was sustained when a further 120 amino acids at the N-terminus of the polypeptide were deleted (DctB5). This protein which has an intact transmitter domain exhibits specific but inefficient phospho-transfer capabilities. Removal of 58 amino acids from the DctB4G C-terminus which included blocks F and G2 of the transmitter domain, rendered the resultant protein (DctB4) incompetent in autophosphorylation. Phosphorylation activity was restored to DctB4 through intramolecular complementation with DctB. Therefore, it would appear that the R. meliloti DctB protein is active as a dimer (or higher order oligomer). Furthermore, the intramolecular complementation experiments indicate that the amino acids 171-291, a predicted periplasmic stretch, play an important role in the dimerization process.
authors.affiliation: Microbiology Department, University College, Cork, Ireland.
authors.forename: L
authors.initials: L
authors.lastname: Giblin
authors.forename: J
authors.initials: J
authors.lastname: Archdeacon
authors.forename: F
authors.initials: F
authors.lastname: O'Gara
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D029382:Dicarboxylic Acid Transporters; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D010766:Phosphorylation; D010957:Plasmids; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D016962:Sinorhizobium meliloti / Q000737:chemistry* / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D029382:Dicarboxylic Acid Transporters; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C073787:DCTB protein, Sinorhizobium; D011494:Protein Kinases