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首页> 外文期刊>Biochemistry >Defining the Bilin Lyase Domain: Lessons from the Extended Phytochrome Superfamily
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Defining the Bilin Lyase Domain: Lessons from the Extended Phytochrome Superfamily

机译：定义Bilin裂解酶域：来自扩展的植物色素超家族的经验教训

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摘要

Through pattern searches of genomic databases,new members of the growing family of phytochrome-related genes were identified and used to construct a 130-180 amino acid motif that delimits the bilin lyase domain,a subdomain of the extended phytochrome family that is sufficient for covalent attachment of linear tetrapyrroles (bilins).To test this hypothesis,portions of locus sll0821,a novel phytochrome-related gene from Synechocystis sp.PCC6803 that encodes a large protein with two potential bilin binding sites,were amplified,and the resombinant apoproteins were tested for bilin binding and phytochromed photoactivity.Our experiments indicated that both sites of this protein,termed Cph2 for cyanobacterial phytochrome 2,possessed bilin lyase activity,resealing two distinct classes of bilin lyase domains-those whose bilin adducts are red,far-red reversible and a second class whose bilin adducts are nonphotochromic.Spectroscopic analysis of photochromic phycocyanobilin and fluorescent phycoerythrobilin adducts of a 24-kDa fragment of Cph2 definitively established that the motif identified by pattern searches represents a bona fide bilin lyase domain.Site-directed mutagenesis of highly conserved charged residues within bilin lyase domains of nearly all members of the extended phytochrome superfamely has identified a glutamate residue critical for bilin binding.
机译：通过基因组数据库的模式搜索，鉴定了正在增长的植物色素相关基因家族的新成员，并将其用于构建130-180个氨基酸基序，该基序限定了Bilin裂解酶结构域，该区域是延伸的植物色素家族中足以共价的子域为了验证该假设，为验证这一假设，对Synechocystis sp.PCC6803的一个新的植物色素相关基因sll0821进行了部分编码，该基因编码具有两个潜在的bilin结合位点的大蛋白，并进行了扩增，并检测了重组重组载脂蛋白我们的实验表明，该蛋白的两个位点，即称为Cph2的蓝细菌植物色素2，都具有bilin裂合酶活性，重新封闭了两类不同的bilin裂合酶结构域，即其bilin加合物是红色的，远红色可逆的和第二类，其胆碱加合物是非光致变色的。光致变色藻蓝蛋白和荧光藻的光谱分析Cph2的24 kDa片段的红霉素加合物确定地确定，通过模式搜索鉴定的基序代表了真正的Bilin裂解酶结构域。几乎所有扩展的植物色素超家族成员的bilin裂解酶结构域内高度保守的带电荷残基的位点诱变鉴定了对Bilin结合至关重要的谷氨酸残基。

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