Synthesis, Properties, and applications of diazotrifluropropanoyl- containing photoactive analogs of farnesyl diphosphate containing modified linkages for enhanced stability

摘要: Photoactive analogs of farnesyl diphosphate (FPP) are useful probes in studies enzymes that employ this molecule as a substrate. Here, we describe the preparation and properties two new FPP contain diazotrifluoropropanoyl photophores linked to geranyl via amide or ester linkages. The amide-linked analog (3) was synthesized 32P-labeled form from geraniol seven steps. Experiments with Saccharomyces cerevisiae protein farnesyltransferase (ScPFTase) showed 3 is an alternative substrate for enzyme. Photolysis experiments [(32)P]3 demonstrate compound labels beta-subunits both geranylgeranyltransferase (types 1 2). However, probe undergoes rearrangement photochemically unreactive isomeric triazolone upon long term storage making it inconvenient use. To address stability issue, ester-linked 4 prepared six steps geraniol. Computational analysis X-ray crystallographic suggest binds transferase (PFTase) similar fashion FPP. Compound also PFTase, selectively photocrosslinks beta-subunit ScPFTase well E. coli farnesyldiphosphate synthase germacrene-producing sesquiterpene Nostoc sp. strain PCC7120 (a cyanobacterial source). Finally, nearly exclusive labeling crude extract observed, suggesting [32P]4 manifests significant selectivity should hence be identifying novel FPP-utilizing preparations.