Kiess M, Hecht H J, Kalisz H M
Department of Molecular Structure Research, Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.
Eur J Biochem. 1998 Feb 15;252(1):90-9. doi: 10.1046/j.1432-1327.1998.2520090.x.
The complete amino acid sequence of glucose oxidase from Penicillium amagasakiense was determined by Edman degradation and mass spectrometry of peptide fragments derived from three different specific proteolytic digests and a cyanogen bromide cleavage. The complete sequence of each monomer comprises 587 amino acid residues, contains three cysteine residues, and seven potential N-glycosylation sites, of which at least five were confirmed to be glycosylated. Glucose oxidase from P. amagasakiense shows a high degree of identity (66%) and 79% similarity to glucose oxidase from Aspergillus niger, and is a member of the glucose-methanol-choline (GMC) oxidoreductase family. The tertiary structures of glucose oxidase from A. niger and cholesterol oxidase from Brevibacterium sterolicum were superimposed to provide a template for the sequence comparison of members of the GMC family. The general topology of the GMC oxidoreductases is conserved, with the exception of the presence of an active site lid in cholesterol oxidase and the insertion of additional structural elements in the substrate-binding domain of alcohol oxidase. The overall structure can be divided into five distinct sequence regions: FAD-binding domain, extended FAD-binding domain, flavin attachment loop and intermediate region, FAD covering lid, and substrate-binding domain. The FAD-binding and the extended FAD-binding domains are composed of several separate sequence regions. The other three regions each comprise a single contiguous sequence. Four major consensus patterns have been identified, including the nucleotide-binding consensus sequence close to their N-termini. The functions of the two motifs recently selected by the Genetics Computer Group, Madison, Wisconsin, as additional signature patterns of the GMC oxidoreductases are discussed. The other consensus patterns belong to either the FAD-binding or the extended FAD-binding domain. In addition, the roles of conserved residues are discussed wherever possible.
通过对源自三种不同特异性蛋白酶消化和溴化氰裂解的肽片段进行埃德曼降解和质谱分析，确定了天草青霉葡萄糖氧化酶的完整氨基酸序列。每个单体的完整序列包含587个氨基酸残基，含有三个半胱氨酸残基和七个潜在的N-糖基化位点，其中至少五个被证实发生了糖基化。天草青霉的葡萄糖氧化酶与黑曲霉的葡萄糖氧化酶具有高度的同一性（66%）和79%的相似性，是葡萄糖-甲醇-胆碱（GMC）氧化还原酶家族的成员。将黑曲霉葡萄糖氧化酶和短杆菌属胆固醇氧化酶的三级结构进行叠加，为GMC家族成员的序列比较提供了模板。GMC氧化还原酶的总体拓扑结构是保守的，除了胆固醇氧化酶中存在活性位点盖子以及醇氧化酶的底物结合结构域中插入了额外的结构元件。整体结构可分为五个不同的序列区域：FAD结合结构域、扩展FAD结合结构域、黄素附着环和中间区域、FAD覆盖盖子以及底物结合结构域。FAD结合结构域和扩展FAD结合结构域由几个单独的序列区域组成。其他三个区域各自包含一个连续的序列。已鉴定出四种主要的共有模式，包括靠近其N端的核苷酸结合共有序列。讨论了最近被威斯康星州麦迪逊市的遗传计算机组选为GMC氧化还原酶额外特征模式的两个基序的功能。其他共有模式属于FAD结合结构域或扩展FAD结合结构域。此外，还尽可能讨论了保守残基的作用。
