title: Catalytic reactions of the homogentisate prenyl transferase involved in plastoquinone-9 biosynthesis.
abstract: Homogentisate solanesyl transferase (HST) catalyzes the prenylation and decarboxylation of homogentisate to form 2-methyl-6-solanesyl-1,4-benzoquinol, the first intermediate in plastoquinone-9 biosynthesis. In vitro, HST from Spinacia oleracea L., Arabidopsis thaliana, and Chlamydomonas reinhardtii were all found to use not only solanesyl diphosphate but also short chain prenyl diphosphates of 10-20 carbon atoms as prenyl donors. Surprisingly, with these donors, prenyl transfer was largely decoupled from decarboxylation, and thus the major products were 6-prenyl-1,4-benzoquinol-2-methylcarboxylates rather than the expected 2-methyl-6-prenyl-1,4-benzoquinols. The 6-prenyl-1,4-benzoquinol-2-methylcarboxylates were not substrates for HST-catalyzed decarboxylation, and the enzyme kinetics associated with forming these products appeared quite distinct from those for 2-methyl-6-prenyl-1,4-benzoquinol formation in respect of catalytic rate, substrate K(m) value, and the pattern of inhibition by haloxydine, a molecule that appeared to act as a dead end mimic of homogentisate. These observations were reconciled into a simple model for the HST mechanism. Here, prenyl diphosphate binds to HST to form at least two alternative complexes that go on to react differently with homogentisate and prenylate it either with or without it first being decarboxylated. It is supposed that solanesyl diphosphate binds tightly and preferentially in the mode that compels prenylation with decarboxylation.
authors.affiliation: Institute for Biology I, Botany, RWTH Aachen University, Worringerweg 1, 52056 Aachen, Germany. rsadre@bio1.rwth-aachen.de
authors.forename: Radin
authors.initials: R
authors.lastname: Sadre
authors.forename: Margrit
authors.initials: M
authors.lastname: Frentzen
authors.forename: Mansoor
authors.initials: M
authors.lastname: Saeed
authors.forename: Tim
authors.initials: T
authors.lastname: Hawkes
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000378:metabolism*; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001671:Biochemistry / Q000379:methods; D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology; D016825:Chlamydomonas reinhardtii / Q000378:metabolism; D002736:Chloroplasts / Q000378:metabolism; D002855:Chromatography, Thin Layer / Q000379:methods; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods; D010971:Plastoquinone / Q000737:chemistry / Q000378:metabolism*; D018724:Spinacia oleracea / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry
chemical_list: D029681:Arabidopsis Proteins; D013729:Terpenes; D019883:Alkyl and Aryl Transferases; C524428:homogentisate solanesyltransferase, Arabidopsis; D010971:Plastoquinone
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