TAILIEUCHUNG - Báo cáo khoa học: Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans

The isoprenoid chain of ubiquinone (Q) is determined bytrans-polyprenyl diphosphate synthase in micro-organisms and presumably in mammals. Because mice and humans produce Q9 and Q10 , they are expected to pos-sess solanesyl and decaprenyl diphosphate synthases as the determining enzyme for a type of ubiquinone. | iFEBS Journal Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans Ryoichi Saiki Ai Nagata Tomohiro Kainou Hideyuki Matsuda and Makoto Kawamukai Faculty of Life and EnvironmentalScience Shimane University Matsue Japan Keywords coenzyme Q isoprenoids prenyl transferase ubiquinone Correspondence Makoto Kawamukai Faculty of Life and EnvironmentalScience Shimane University 1060 Nishikawatsu Matsue 690-8504 Japan Fax 81 852 32 6092 Tel 81 852 32 6587 E-mail kawamuka@ Received 12 July 2005 revised 23 August 2005 accepted 5 September 2005 doi The isoprenoid chain of ubiquinone Q is determined by trans-polyprenyl diphosphate synthase in micro-organisms and presumably in mammals. Because mice and humans produce Q9 and Q10 they are expected to possess solanesyl and decaprenyl diphosphate synthases as the determining enzyme for a type of ubiquinone. Here we show that murine and human solanesyl and decaprenyl diphosphate synthases are heterotetramers composed of newly characterized hDPS1 mSPS1 and hDLP1 mDLP1 which have been identified as orthologs of Schizosaccharomyces pombe Dps1 and Dlp1 respectively. Whereas hDPSl or mSPSl can complement the S. po-mbe dpsl disruptant neither hDLPl nor mDLPl could complement the S. pombe dLpl disruptant. Thus only hDPS1 and mSPS1 are functional orthologs of SpDps1. Escherichia coli was engineered to express murine and human SpDps1 and or SpDlp1 homologs and their ubiquinone types were determined. Whereas transformants expressing a single component produced only Q8 of E. coli origin double transformants expressing mSPS1 and mDLP1 or hDPS1 and hDLP1 produced Q9 or Q10 respectively and an in vitro activity of solanesyl or decaprenyl diphosphate synthase was verified. The complex size of the human and murine long-chain transprenyl diphosphate synthases as estimated by gel-filtration chromatography indicates that they consist of heterotetramers. Expression in .

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