家蚕昆虫芳基烷基胺N-乙酰基转移酶的克隆与特性分析
Cloning and characterization of insect arylalkylamine N-acetyltransferase from Bombyx mori.
作者信息
Tsugehara Taketo, Iwai Sachio, Fujiwara Yoshihiro, Mita Kazuei, Takeda Makio
机构信息
Division of Molecular Science, Graduate School of Science and Technology, Kobe University, Kobe, Hyogo, Japan.
出版信息
Comp Biochem Physiol B Biochem Mol Biol. 2007 Jul;147(3):358-66. doi: 10.1016/j.cbpb.2006.10.112. Epub 2007 Feb 7.
Arylalkylamine N-acetyltransferase (AANAT) catalyzes N-acetylation of arylarkylamines. A cDNA of Bombyx mori insect AANAT (Bm-iAANAT) was found by searching an expressed-sequence tag (EST) database of B. mori (SilkBase). The cDNA encoded a 261 amino acid protein. The mRNA of Bm-iAANAT was expressed in eggs, larvae, adults and various tissues. Recombinant Bm-iAANAT protein was expressed in Sf9 cells by a baculovirus expression system. The AANAT activity of Bm-iAANAT was inhibited by high concentrations (over 0.01 mM) of tryptamine used as a substrate. The Bm-iAANAT acetylated tryptamine, serotonin, dopamine, octopamine, tyramine and norepinephrine. This is the first report of a cloned AANAT that acetylated norepinephrine. These results suggest that Bm-iAANAT is a novel member of insect AANAT family with unique kinetic properties and a broad substrate range.
芳基烷基胺N - 乙酰基转移酶（AANAT）催化芳基烷基胺的N - 乙酰化反应。通过搜索家蚕（SilkBase）的表达序列标签（EST）数据库，发现了家蚕昆虫AANAT（Bm - iAANAT）的cDNA。该cDNA编码一个261个氨基酸的蛋白质。Bm - iAANAT的mRNA在卵、幼虫、成虫及各种组织中表达。重组Bm - iAANAT蛋白通过杆状病毒表达系统在Sf9细胞中表达。用作底物的高浓度（超过0.01 mM）色胺抑制了Bm - iAANAT的AANAT活性。Bm - iAANAT使色胺、5 - 羟色胺、多巴胺、章鱼胺、酪胺和去甲肾上腺素乙酰化。这是关于克隆的AANAT使去甲肾上腺素乙酰化的首次报道。这些结果表明Bm - iAANAT是昆虫AANAT家族的一个新成员，具有独特的动力学特性和广泛的底物范围。
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