Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans
Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans
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DOI：
10.1111/j.1742-4658.2005.04956.x
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发表时间：
2005-11-01
期刊：
影响因子：
5.4
通讯作者：
Kawamukai, M
中科院分区：
文献类型：
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作者：
Saiki, R;Nagata, A;Kawamukai, M
泛醌（Q）的类异戊二烯链由微生物和哺乳动物中的反式聚异戊二烯基二磷酸合酶决定。因为小鼠和人类产生Q（9）和Q（10），所以预期它们具有茄呢基和十异戊二烯基二磷酸脱氢酶作为泛醌类型的决定酶。在这里，我们表明，鼠和人的茄呢基和decaprenyl二磷酸脱氢酶的异源四聚体组成的新特征的hDPS 1（mSPS 1）和hDLP 1（mDLP 1），已被确定为直系同源的裂殖酵母菌粟酒裂殖酵母DPS 1和DLP 1，分别。而hDPS 1或mSPS 1可以与S. pombe dps 1破坏剂，hDLP 1和mDLP 1均不能与S.粟酒裂殖酵母dLp 1干扰子。因此，只有hDPS 1和mSPS 1是SpDps 1的功能性直系同源物。大肠杆菌工程表达鼠和人SpDps 1和/或SpDlp 1同源物，并确定其泛醌类型。而表达单一组分的转化子仅产生E. coli来源，表达mSPS 1和mDLP 1或hDPS 1和hDLP 1的双转化体分别产生Q（9）或Q（10），并验证了茄呢基或十异戊二烯基二磷酸合酶的体外活性。人和鼠的长链trans-prenyl二磷酸脱氢酶的复杂的大小，估计通过凝胶过滤色谱法，表明它们由异四聚体。在大肠杆菌中的表达大肠杆菌的异源组合，即mSPS 1和hDLP 1或hDPS 1和mDLP 1，产生了Q（9）和Q（10），表明这两种组分都参与决定泛醌侧链。因此，我们确定了哺乳动物中决定泛醌侧链的酶的组分，它们类似于S。粟酒，但不是植物或酿酒酵母，酶的类型。
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.