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首页> 外文期刊>The FEBS journal >Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans
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Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans

机译：小鼠和人类中茄基和癸二烯基二磷酸合成酶的表征

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摘要

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 Q(9) and Q(10), 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 hDPS1 or mSPS1 can complement the S. pombe dps1 disruptant, neither hDLP1 nor mDLP1 could complement the S. pombe dLp1 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 Q(8) of E. coli origin, double transformants expressing mSPS1 and mDLP1 or hDPS1 and hDLP1 produced Q(9) or Q(10), respectively, and an in vitro activity of solanesyl or decaprenyl diphosphate synthase was verified. The complex size of the human and murine long-chain trans-prenyl diphosphate synthases, as estimated by gel-filtration chromatography, indicates that they consist of heterotetramers. Expression in E. coli of heterologous combinations, namely, mSPS1 and hDLP1 or hDPS1 and mDLP1, generated both Q(9) and Q(10), indicating both components are involved in determining the ubiquinone side chain. Thus, we identified the components of the enzymes that determine the side chain of ubiquinone in mammals and they resembles the S. pombe, but not plant or Saccharomyces cerevisiae, type of enzyme.
机译：泛醌（Q）的类异戊二烯链是由反式聚异戊二烯二磷酸合酶在微生物中（大概在哺乳动物中）确定的。由于小鼠和人类会产生Q（9）和Q（10），因此预期它们具有茄基和癸二烯基二磷酸合酶作为一种泛醌的决定酶。在这里，我们显示鼠和人茄基和癸二烯基二磷酸合酶是由新鉴定的hDPS1（mSPS1）和hDLP1（mDLP1）组成的异四聚体，它们分别被鉴定为粟酒裂殖酵母Dps1和Dlp1的直系同源物。 hDPS1或mSPS1可以补充粟酒裂殖酵母dps1破坏物，而hDLP1和mDLP1都不能补充粟酒裂殖酵母dLp1破坏物。因此，只有hDPS1和mSPS1是SpDps1的功能直系同源物。大肠杆菌经过工程改造可以表达鼠和人SpDps1和/或SpDlp1同源物，并确定了它们的泛醌类型。表达单个成分的转化子仅产生Q.8大肠杆菌，表达mSPS1和mDLP1或hDPS1和hDLP1的双重转化子分别产生Q（9）或Q（10），而茄基或癸二烯基的体外活性验证了二磷酸合酶。通过凝胶过滤色谱估计的人和鼠长链反式异戊二烯基二磷酸合酶的复杂大小表明，它们由异四聚体组成。异源组合（即mSPS1和hDLP1或hDPS1和mDLP1）在大肠杆菌中的表达产生了Q（9）和Q（10），表明这两个成分都参与确定泛醌侧链。因此，我们鉴定了确定哺乳动物中泛醌侧链的酶成分，它们类似于粟酒裂殖酵母，而不是植物或酿酒酵母。

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