The Establishment and Application of Anti-Pseudomonas aeruginosa Cell-based Screening Model Targeting to SecA

Abstract:

The most of secreted proteins are exported by Sec translocase (secretion pathway). SecA ATPase is one of the most important subunit in the Sec translocase, which is preprotein translocase nanomotor that undergo membrane insertion and deinsertion to drive preprotein across the bacterial inner membrane, and SecA is indispensable to bacteria. It should be presumed that the compound which inhibits the activity of SecA ATPase probably can be used as the candidate of bactericide. A secA gene from Pseudomonas aeruginosa PAO1 was amplified and expressed in Escherichia coli BL21.19 (secA13). It has been shown that the wild-type SecA of Pseudomonas aeruginosa could fully complement the E. coli amber (secA13) mutant at the non-permissive temperature. So a cell level screening model targeting on SecA was established based on the above result. The inhibition of PaSecA ATPase activity was applied to validate the specificity of the cell-based method. Two positive samples based on both of cell and enzyme activities will be further studied.