Human fatty acid ω-hydroxylase, CYP4A11:: determination of complete genomic sequence and characterization of purified recombinant protein
Human fatty acid ω-hydroxylase, CYP4A11:: determination of complete genomic sequence and characterization of purified recombinant protein
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DOI：
10.1006/abbi.2000.1831
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发表时间：
2000-06-15
影响因子：
3.9
通讯作者：
Kishimoto, T
中科院分区：
文献类型：
--
作者：
Kawashima, H;Naganuma, T;Kishimoto, T
人类脂肪酸omega-羟化酶基因CYP4A11已从人类BAC文库中分离出来，并确定了其完整的基因组序列。 CYP4A11 基因全长 12,568 bp，包含 12 个外显子，在 CYP4A11 基因的 5' 侧翼区域未观察到据报道参与氯贝酸诱导 CYP4A6 的已知 PPAR 识别元件 (PPRE)。 使用 pCWOri 表达载体在大肠杆菌中表达的重组 CYP4A11 蛋白被纯化几乎电泳均质的状态，具体含量为 6.4 nmol P450/mg 蛋白质。该 P450 对月桂酸酯表现出 omega-羟基化活性，P450 的周转数为 14.7 nmol/min/nmol。 P450的表观K-m和V-max值分别为56.7μM和15.2nmol/min/nmol。它还显示出对棕榈酸酯的 omega-羟基化活性，P450 的周转数为 0.78 nmol/min/nmol。尽管其他小组的几份报告描述 CYP4A11 制剂催化花生四烯酸的 omega-羟基化，但我们纯化的重组蛋白对花生四烯酸和前列腺素 A (1) 没有表现出活性。 (C) 2000 年学术出版社。
The gene of the human fatty acid omega-hydroxylase, CYP4A11, has been isolated from a human BAC library, and its complete genomic sequence has been determined. The CYP4A11 gene spanned 12,568 bp and contained 12 exons, The known PPAR recognition elements (PPRE), which were reported to be involved in the induction of CYP4A6 by clofibric acid, were not observed within the 5'-flanking region of the CYP4A11 gene, The recombinant CYP4A11 protein expressed in Escherichia coli using the pCWOri expression vector was purified to an almost electrophoretically homogeneous state with a specific content of 6.4 nmol of P450/mg of protein. This P450 exhibited omega-hydroxylation activity toward laurate, with a turnover number of 14.7 nmol/min/nmol of P450. The apparent K-m and V-max values were 56.7 mu M and 15.2 nmol/min/nmol of P450, respectively. It also showed omega-hydroxylation activity toward palmitate, with a turnover number of 0.78 nmol/min/nmol of P450, Although several reports from other groups described that CYP4A11 preparations catalyzed omega-hydroxylation of arachidonic acid, our purified recombinant protein exhibited no activity toward arachidonic acid nor prostaglandin A(1). (C) 2000 Academic Press.