Basic Local Alignment Search Tool studies

The first bacterial DPCK to be cloned and characterized was the E. coli CoaE protein coaE gene product, previously yacE), which was identified by a basic local alignment search tool (BLAST) using the N-terminal sequence of the wild-type enzyme purified from C. ammoniagenes The enzyme is a 22.6-kDa monomer in solution and exhibits apparent values of 140 pmol 1 and 740 pmol for ATP and dephospho-CoA, respectively. It displays poor activity with adenosine, AMP, and adenosine phosphosulfate (APS) as alternate phosphoryl acceptors, which all shows less than 8% activity compared with the natural substrate. Studies on the bifiinctional PPAT/ DPCK protein show that the normally reversible PPAT activity becomes irreversible when it is coupled to DPCK, most probably because the latter activity demonstrates a low K- for dephospho-CoA of 5.2 L5 pmol 1 (its for ATP was found to be 192 14 pmol 1 ). ° Similar values were determined in an independent study. ... 

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