Catalase-Related Allene Oxide Synthase, on a Biosynthetic Route to Fatty Acid Cyclopentenones: Expression and Assay of the Enzyme and Preparation of the 8R-HPETE Substrate.
Catalase-Related Allene Oxide Synthase, on a Biosynthetic Route to Fatty Acid Cyclopentenones: Expression and Assay of the Enzyme and Preparation of the 8R-HPETE Substrate.
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DOI：
10.1016/bs.mie.2018.02.019
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发表时间：
2018
影响因子：
--
通讯作者：
Brash AR
中科院分区：
文献类型：
--
作者：
Brash AR
过氧化氢酶相关的丙二烯氧化物合成酶(CAOS)是一种血红素蛋白，能以每秒1000左右的周转速度将特定的脂肪酸氢过氧化氢转化为不稳定的丙二烯氧化物中间体。脂肪酸丙二烯氧化物是海洋体系中形成环戊烯酮或水解物的中间体，最著名的是与前列腺素相关的棒形酮。虽然CAOS活性部位周围的关键催化氨基酸残基与真正的过氧化氢酶相同，但CAOS不与过氧化氢反应。CAOS仅作为天然存在的融合蛋白的N-末端结构域与提供氢过氧化氢底物的C-末端脂氧合酶(LOX)结构域一起出现。在海洋无脊椎动物中，8R-LOX结构域将花生四烯酸转化为8R-羟基过氧乙烯-二十碳四烯酸(8R-HPETE)，而CAOS结构域形成8，9-环氧丙二烯氧化物。海鞭八爪珊瑚Plexaura homomalla的融合蛋白是具有单个结构域晶体结构的原型模型。CAOS(43kD)在大肠杆菌中有很好的表达，产量高达100 mg/L。本文详细介绍了同型假单胞菌CAOS的表达和活性测定及其8R-HPETE底物的两种制备方法。这本书中的另一篇文章集中在P.homomalla 8R-LOX上。
Catalase-related allene oxide synthase (cAOS) is a hemoprotein that converts a specific fatty acid hydroperoxide to an unstable allene oxide intermediate at turnover rates in the order of 1000 per second. Fatty acid allene oxides are intermediates in the formation of cyclopentenone or hydrolytic products in marine systems, most notably the prostanoid-related clavulones. Although the key catalytic amino acid residues around the active site of cAOS are the same as in true catalases, cAOS does not react with hydrogen peroxide. cAOS occur exclusively as the N-terminal domain of a naturally occurring fusion protein with a C-terminal lipoxygenase (LOX) domain that supplies the hydroperoxide substrate. In marine invertebrates, an 8R-LOX domain converts arachidonic acid to 8R-hydroperoxy-eicosatetraenoic acid (8R-HPETE) and the cAOS domain forms an 8,9-epoxy allene oxide. The fusion protein from the sea-whip octocoral Plexaura homomalla is the prototypical model with crystal structures of the individual domains. The cAOS (43 kD) expresses exceptionally well in E. coli, with yields of up to 100 mg/L. This article describes in detail expression and assay of the P. homomalla cAOS and two methods for the preparation of its 8R-HPETE substrate. Another article in this volume focuses on the P. homomalla 8R-LOX.
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影响因子：
4.8
作者：
Boeglin, William E.;Brash, Alan R.
通讯作者：
Brash, Alan R.
影响因子：
1.6
作者：
HOFFMAN, BJ;BROADWATER, JA;KENEALY, WR
通讯作者：
KENEALY, WR
影响因子：
1.9
作者：
Teder T;Boeglin WE;Brash AR
通讯作者：
Brash AR
影响因子：
2.9
作者：
Gilbert, Nathaniel C.;Niebuhr, Marc;Newcomer, Marcia E.
通讯作者：
Newcomer, Marcia E.
DOI：
10.1107/s1744309107050993
发表时间：
2007-11-01
影响因子：
0.9
作者：
Neau, David B.;Gilbert, Nathaniel C.;Newcomer, Marcia E.
通讯作者：
Newcomer, Marcia E.