title: A photoactive isoprenoid diphosphate analogue containing a stable phosphonate linkage: synthesis and biochemical studies with prenyltransferases.
abstract: A number of biochemical processes rely on isoprenoids, including the post-translational modification of signaling proteins and the biosynthesis of a wide array of compounds. Photoactivatable analogues have been developed to study isoprenoid utilizing enzymes such as the isoprenoid synthases and prenyltransferases. While these initial analogues proved to be excellent structural analogues with good cross-linking capability, they lack the stability needed when the goals include isolation of cross-linked species, tryptic digestion, and subsequent peptide sequencing. Here, the synthesis of a benzophenone-based farnesyl diphosphate analogue containing a stable phosphonophosphate group is described. Inhibition kinetics, photolabeling experiments, as well as X-ray crystallographic analysis with a protein prenyltransferase are described, verifying this compound as a good isoprenoid mimetic. In addition, the utility of this new analogue was explored by using it to photoaffinity label crude protein extracts obtained from Hevea brasiliensis latex. Those experiments suggest that a small protein, rubber elongation factor, interacts directly with farnesyl diphosphate during rubber biosynthesis. These results indicate that this benzophenone-based isoprenoid analogue will be useful for identifying enzymes that utilize farnesyl diphosphate as a substrate.
authors.affiliation: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, USA.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: DeGraw
authors.forename: Zongbao
authors.initials: Z
authors.lastname: Zhao
authors.forename: Corey L
authors.initials: CL
authors.lastname: Strickland
authors.forename: A Huma
authors.initials: AH
authors.lastname: Taban
authors.forename: John
authors.initials: J
authors.lastname: Hsieh
authors.forename: Michael
authors.initials: M
authors.lastname: Jefferies
authors.forename: Wenshuang
authors.initials: W
authors.lastname: Xie
authors.forename: David K
authors.initials: DK
authors.lastname: Shintani
authors.forename: Colleen M
authors.initials: CM
authors.lastname: McMahan
authors.forename: Katrina
authors.initials: K
authors.lastname: Cornish
authors.forename: Mark D
authors.initials: MD
authors.lastname: Distefano
mesh_terms: D001577:Benzophenones / Q000737:chemistry; D002384:Catalysis; D003432:Cross-Linking Reagents / Q000737:chemistry; D018360:Crystallography, X-Ray; D004122:Dimethylallyltranstransferase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D028482:Hevea / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D013058:Mass Spectrometry; D008958:Models, Molecular; D015394:Molecular Structure; D063065:Organophosphonates / Q000737:chemistry*; D010777:Photochemistry; D011106:Polyisoprenyl Phosphates / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D001577:Benzophenones; D003432:Cross-Linking Reagents; D004791:Enzyme Inhibitors; D063065:Organophosphonates; D011106:Polyisoprenyl Phosphates; C047723:benzophenone; D004122:Dimethylallyltranstransferase