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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1
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Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1

机译:使用互补的阳离子和阴离子重原子盐衍生物的结构来解决细胞色素P450 46 a1

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摘要

Human cytochrome P450 46A1 (CYP46A1) is one of the key enzymes in cholesterol homeostasis in the brain. The crystallization and heavy-atom structure solution of an active truncated CYP46A1 in complex with the high-affinity substrate analogue cholesterol-3-sulfate (CH-3S) is reported. The 2.6 A structure of CYP46A1-CH-3S was solved using both anion and cation heavy-atom salts. In addition to the native anomalous signal from the haem iron, an NaI anion halide salt derivative and a complementary CsCl alkali-metal cation salt derivative were used. The general implications of the use of halide and alkali-metal quick soaks are discussed. The importance of using isoionic strength buffers, the titration of heavy-atom salts into different ionic species and the role of concentration are considered. It was observed that cation/anion-binding sites will occasionally overlap, which could negatively impact upon mixed RbBr soaks used for multiple anomalous scatterer MAD (MMAD). The use of complementary cation and anion heavy-atom salt derivatives is a convenient and powerful tool for MIR(AS) structure solution.
机译:人类细胞色素P450 46 a1 (CYP46A1)是其中一个关键酶在胆固醇体内平衡大脑。一个活跃的截断CYP46A1的结构方案在复杂的高亲和性衬底模拟cholesterol-3-sulfate (CH-3S)报道。解决了使用这两种阴离子和阳离子重原子盐。血红素铁,奈阴离子卤盐导数和互补中海碱金属阳离子盐衍生物。影响使用的卤化物讨论了碱金属快速浸泡。使用等离子强度缓冲的重要性,重原子盐滴定法的不同离子种类和浓度的作用考虑。阳离子/阴离子结合网站偶尔会在混合的重叠,这将产生负面影响RbBr浸泡用于多个异常散射体疯了(MMAD)。阴离子盐重原子衍生物是一种方便的和米尔()结构解决方案的有力工具。

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