A versatile photoactivatable probe designed to label the diphosphate binding site of farnesyl diphosphate utilizing enzymes

Olivier Henry, Fernando Lopez-Gallego, Sean A. Agger, Claudia Schmidt-Dannert, Stephanie Sen, David Shintani, Katrina Cornish, Mark D. Distefano

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Farnesyl diphosphate (FPP) is a substrate for a diverse number of enzymes found in nature. Photoactive analogues of isoprenoid diphosphates containing either benzophenone, diazotrifluoropropionate or azide groups have been useful for studying both the enzymes that synthesize FPP as well as those that employ FPP as a substrate. Here we describe the synthesis and properties of a new class of FPP analogues that links an unmodified farnesyl group to a diphosphate mimic containing a photoactive benzophenone moiety; thus, importantly, these compounds are photoactive FPP analogues that contain no modifications of the isoprenoid portion of the molecule that may interfere with substrate binding in the active site of an FPP utilizing enzyme. Two isomeric compounds containing meta- and para-substituted benzophenones were prepared. These two analogues inhibit Saccharomyces cerevisiae protein farnesyltransferase (ScPFTase) with IC50 values of 5.8 (meta isomer) and 3.0 μM (para isomer); the more potent analogue, the para isomer, was shown to be a competitive inhibitor of ScPFTase with respect to FPP with a KI of 0.46 μM. Radiolabeled forms of both analogues selectively labeled the β-subunit of ScPFTase. The para isomer was also shown to label Escherichia coli farnesyl diphosphate synthase and Drosophila melanogaster farnesyl diphosphate synthase. Finally, the para isomer was shown to be an alternative substrate for a sesquiterpene synthase from Nostoc sp. strain PCC7120, a cyanobacterial source; the compound also labeled the purified enzyme upon photolysis. Taken together, these results using a number of enzymes demonstrate that this new class of probes should be useful for a plethora of studies of FPP-utilizing enzymes.

Original languageEnglish (US)
Pages (from-to)4797-4805
Number of pages9
JournalBioorganic and Medicinal Chemistry
Volume17
Issue number13
DOIs
StatePublished - Jul 1 2009

Bibliographical note

Funding Information:
This research was supported by the National Institutes of Health Grant GM58442 (M.D.D.), the National Science Foundation Grant DBI 0321690 (D.S.) and funds from Yulex Corp.

Keywords

  • Benzophenone
  • Farnesyl diphosphate synthase
  • Farnesyltransferase
  • Photoaffinity labeling
  • Sesquiterpene cyclase

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