The conserved baculovirus protein p33 (Ac92) is a flavin adenine dinucleotide-linked sulfhydryl oxidase
The conserved baculovirus protein p33 (Ac92) is a flavin adenine dinucleotide-linked sulfhydryl oxidase
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DOI：
10.1016/j.virol.2009.04.006
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发表时间：
2009-06-05
期刊：
影响因子：
3.7
通讯作者：
Merrill, G. F.
中科院分区：
文献类型：
--
作者：
Long, C. M.;Rohrmann, G. F.;Merrill, G. F.
苜蓿银纹夜蛾杆状病毒的开放阅读框92（Ac 92）是存在于所有测序的杆状病毒基因组中的约30个核心基因之一。计算机分析预测，Ac 92编码的蛋白质（称为p33）和它的几个杆状病毒直系同源物与黄素腺嘌呤二核苷酸（FAD）连接的巯基氧化酶家族有关。这些蛋白质的比对表明，虽然它们是高度多样化的，但存在一些与Erv 1 p/Alrp家族的巯基氧化酶共同的氨基酸。预测这些保守氨基酸中的一些与FAD的异咯嗪和腺嘌呤组分堆叠，而其他的参与电子转移。为了研究这种关系，Ac 92在细菌中表达为His-标记的融合蛋白，纯化，并通过荧光光度法和其酶活性进行表征。发现纯化蛋白的颜色（黄色）和吸收光谱与其为含FAD蛋白一致。此外，使用二硫苏糖醇和硫氧还蛋白作为底物证明其具有巯基氧化酶活性。(c)2009 Elsevier Inc. All rights reserved.
Open reading frame 92 of the Autographa californica baculovirus (Ac92) is one of about 30 core genes present in all sequenced baculovirus genomes. Computer analyses predicted that the Ac92 encoded protein (called p33) and several of its baculovirus orthologs were related to a family of flavin adenine dinucleotide (FAD)-linked sulfhydryl oxidases. Alignment of these proteins indicated that, although they were highly diverse, a number of amino acids in common with the Erv1p/Alrp family of sulfhydryl oxidases are present. Some of these conserved amino acids are predicted to stack against the isoalloxazine and adenine components of FAD, whereas others are involved in electron transfer. To investigate this relationship, Ac92 was expressed in bacteria as a His-tagged fusion protein, Purified, and characterized both spectrophotometrically and for its enzymatic activity. The Purified protein was found to have the color (yellow) and absorption spectrum consistent with it being a FAD-containing protein. Furthermore, it was demonstrated to have sulfhydryl oxidase activity using dithiothreitol and thioredoxin as Substrates. (c) 2009 Elsevier Inc. All rights reserved.