Chemotaxis in Escherichia coli: associations of protein components.
Interactions between protein components of the chemotaxis mechanism in Escherichia coli were investigated by using the cleavable cross-linking reagent, dithiobis(succinimidyl propionate). Two methods were used to allow detection of chemotaxis-specific proteins in intact cells. The first method was to program their synthesis in the presence of [35S]methionine using lambda E. coli hybrid phages which carry the chemotaxis genes. The second method was to label endogenous methyl-accepting chemotaxis proteins (MCP's), with the methyl donor S-adenosyl-L-[methyl-3H]methionine, after permeabilizing the cells with EGTA. Physical associations between proteins were analyzed, after cross-linking, by two dimensional NaDodSO4-polyacrylamide gel electrophoresis. Both labeling methods demonstrate that MCP I and MCP II exist as functional tetramers. Other proteins involved with chemotaxis were found to form dimers and higher polymers. Phage-directed products of cheW, cheX, motA, and cheA formed dimers. CheB and hag products formed multimers. A number of apparent interactions between different gene products were detected as well. Products of cheB, cheW, cheZ, motA, and motB were found to form complexes with other gene products. Included are results consistent with interactions between the products of cheB and cheZ.
D001426:Bacterial Proteins / Q000378:metabolism*; D010582:Bacteriophage lambda / Q000378:metabolism; D002633:Chemotaxis; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004220:Disulfides / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005796:Genes; D006824:Hybridization, Genetic; D013388:Succinimides / Q000494:pharmacology*
D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D004220:Disulfides; D013388:Succinimides; C011240:dithiobis(succinimidylpropionate)