title: Vitamin E biosynthesis: functional characterization of the monocot homogentisate geranylgeranyl transferase.
abstract: The biosynthesis of the tocotrienol and tocopherol forms of vitamin E is initiated by prenylation of homogentisate. Geranylgeranyl diphosphate (GGDP) is the prenyl donor for tocotrienol synthesis, whereas phytyl diphosphate (PDP) is the prenyl donor for tocopherol synthesis. We have previously shown that tocotrienol synthesis is initiated in monocot seeds by homogentisate geranylgeranyl transferase (HGGT). This enzyme is related to homogentisate phytyltransferase (HPT), which catalyzes the prenylation step in tocopherol synthesis. Here we show that monocot HGGT is localized in the plastid and expressed primarily in seed endosperm. Despite the close structural relationship of monocot HGGT and HPT, these enzymes were found to have distinct substrate specificities. Barley (Hordeum vulgare cv. Morex) HGGT expressed in insect cells was six times more active with GGDP than with PDP, whereas the Arabidopsis HPT was nine times more active with PDP than with GGDP. However, only small differences were detected in the apparent Km values of barley HGGT for GGDP and PDP. Consistent with its in vitro substrate properties, barley HGGT generated a mixture of tocotrienols and tocopherols when expressed in the vitamin E-null vte2-1 mutant lacking a functional HPT. Relative levels of tocotrienols and tocopherols produced in vte2-1 differed between organs and growth stages, reflective of the composition of plastidic pools of GGDP and PDP. In addition, HGGT was able to functionally substitute for HPT to rescue vte2-1-associated phenotypes, including reduced seed viability and increased fatty acid oxidation of seed lipids. Overall, we show that monocot HGGT is biochemically distinct from HPT, but can replace HPT in important vitamin E-related physiological processes.
authors.affiliation: Donald Danforth Plant Science Center, 975 North Warson Road, Saint Louis, MO 63132, USA.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Yang
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Cahoon
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Hunter
authors.forename: Chunyu
authors.initials: C
authors.lastname: Zhang
authors.forename: Jixiang
authors.initials: J
authors.lastname: Han
authors.forename: Trissa
authors.initials: T
authors.lastname: Borgschulte
authors.forename: Edgar B
authors.initials: EB
authors.lastname: Cahoon
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D003080:Cold Temperature; D056625:Endosperm / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D001467:Hordeum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism; D018087:Plastids / Q000378:metabolism; D054876:Prenylation; D018411:Spodoptera / Q000201:enzymology / Q000378:metabolism; D013379:Substrate Specificity; D024505:Tocopherols / Q000032:analysis / Q000378:metabolism; D024508:Tocotrienols / Q000032:analysis / Q000378:metabolism; D014908:Triticum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D014810:Vitamin E / Q000096:biosynthesis*
chemical_list: D029681:Arabidopsis Proteins; D010940:Plant Proteins; D024508:Tocotrienols; D014810:Vitamin E; D019883:Alkyl and Aryl Transferases; C446531:HPT1 protein, Arabidopsis; C569419:homogentisate geranylgeranyl transferase, Hordeum vulgare; D024505:Tocopherols