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Synthesis, properties, and applications of diazotrifluropropanoyl-containing photoactive analogs of farnesyl diphosphate containing modified linkages for enhanced stability.
- Source :
-
Chemical biology & drug design [Chem Biol Drug Des] 2010 Jan; Vol. 75 (1), pp. 51-67. - Publication Year :
- 2010
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Abstract
- Photoactive analogs of farnesyl diphosphate (FPP) are useful probes in studies of enzymes that employ this molecule as a substrate. Here, we describe the preparation and properties of two new FPP analogs that contain diazotrifluoropropanoyl photophores linked to geranyl diphosphate via amide or ester linkages. The amide-linked analog (3) was synthesized in 32P-labeled form from geraniol in seven steps. Experiments with Saccharomyces cerevisiae protein farnesyltransferase (ScPFTase) showed that 3 is an alternative substrate for the enzyme. Photolysis experiments with [(32)P]3 demonstrate that this compound labels the beta-subunits of both farnesyltransferase and geranylgeranyltransferase (types 1 and 2). However, the amide-linked probe 3 undergoes a rearrangement to a photochemically unreactive isomeric triazolone upon long term storage making it inconvenient to use. To address this stability issue, the ester-linked analog 4 was prepared in six steps from geraniol. Computational analysis and X-ray crystallographic studies suggest that 4 binds to protein farnesyl transferase (PFTase) in a similar fashion as FPP. Compound 4 is also an alternative substrate for PFTase, and a 32P-labeled form selectively photocrosslinks the beta-subunit of ScPFTase as well as E. coli farnesyldiphosphate synthase and a germacrene-producing sesquiterpene synthase from Nostoc sp. strain PCC7120 (a cyanobacterial source). Finally, nearly exclusive labeling of ScPFTase in crude E. coli extract was observed, suggesting that [32P]4 manifests significant selectivity and should hence be useful for identifying novel FPP-utilizing enzymes in crude protein preparations.
- Subjects :
- Binding Sites
Farnesyltranstransferase metabolism
Kinetics
Photoaffinity Labels
Polyisoprenyl Phosphates chemistry
Polyisoprenyl Phosphates pharmacology
Saccharomyces cerevisiae Proteins chemistry
Sesquiterpenes chemistry
Sesquiterpenes pharmacology
Structure-Activity Relationship
Substrate Specificity
Polyisoprenyl Phosphates chemical synthesis
Sesquiterpenes chemical synthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1747-0285
- Volume :
- 75
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Chemical biology & drug design
- Publication Type :
- Academic Journal
- Accession number :
- 19954434
- Full Text :
- https://doi.org/10.1111/j.1747-0285.2009.00914.x