A mechanism-based fluorescence transfer assay for examining ketosynthase selectivity.

ABSTRACT: Since their discovery, polyketide synthases have received massive attention from researchers hoping to harness their potential as a platform for generating new and improved therapeutics. Despite significant strides toward this end, inherent specificities within the enzymes responsible for polyketide production have severely limited these efforts. We have developed a mechanism-based, fluorescence transfer assay for a key enzyme component of all polyketide synthases, the ketosynthase domain. As demonstrated, this method can be used with both ketosynthase-containing didomains and full modules. As proof of principle, the ketosynthase domain from module 6 of the 6-deoxyerythronolide synthase is examined for its ability to accept a variety of simple thioester substrates. Consistent with its natu

SUBMITTER: Prasad G 

PROVIDER: S-EPMC4232300 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature