Phosphorylation assays of chemotaxis two-component system proteins in Borrelia burgdorferi.

摘要: Abstract Borrelia burgdorferi has a complex chemotaxis signal transduction system with multiple gene homologs similar to those found in Escherichia coli and Bacillus subtilis . The B. genome sequence encodes two cheA , three cheY cheW cheB cheR but no cheZ genes. Instead of cheZ, contains different CheY‐P phosphatase, referred as cheX histidine kinases (CheA1 CheA2) response regulators (CheY1, CheY2, CheY3) possess all the domains functional residues E. CheA CheY, respectively. Understanding protein phosphorylation is critical unraveling many biological processes, including transduction, motility, growth control, metabolism, disease processes. coli, Salmonella enterica serovar Typhimurium, systems have been studied extensively, providing models understand signaling Lyme spirochete Both genetic approaches biochemical analyses are essential understanding its two‐component systems. Specifically, inactivation studies assess importance specific genes motility under certain conditions. Furthermore, help determine following vitro reactions: (1) extent that kinases, CheA1 CheA2, autophosphorylated using ATP; (2) transfer phosphate from CheA1‐P CheA2‐P each CheY species; (3) dephosphorylation species by CheX. We hypothesize characterizing will facilitate how periplasmic flagellar bundles located near end cells coordinately regulated for chemotaxis. During chemotaxis, these bacteria run, pause (stop/flex), reverse (run again). This chapter describes protocols assessing autophosphorylation, CheA‐P dephosphorylation.