克隆的枯草芽孢杆菌谷氨酰胺phosphoribosylpyrophosphate amidotransferase基因在大肠杆菌。plasmid-encoded酶的核苷酸序列测定和属性。
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Makaroff CA, Zalkin H,瑞士人RL, Vollmer SJ
克隆的枯草芽孢杆菌谷氨酰胺phosphoribosylpyrophosphate amidotransferase基因在大肠杆菌。plasmid-encoded酶的核苷酸序列测定和属性。
J生物化学杂志。1983年9月10日,258 (17):10586 - 93。
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6411717 (在PubMed]
- 文摘
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枯草芽孢杆菌基因编码谷氨酰胺phosphoribosylpyrophosphate amidotransferase (amidophosphoribosyltransferase)在pBR322克隆。这个基因被指定purF类比与相应基因在大肠杆菌。枯草芽孢杆菌purF从质粒在大肠杆菌表达的启动子。plasmid-encoded酶是体内功能,补充一个大肠杆菌purF突变株。的核苷酸序列1651 -碱基对枯草芽孢杆菌DNA片段确定,因此本地化1428 -碱基对结构基因。476个氨基酸残基的主要翻译产品推导了从DNA序列。与先前决定NH2-terminal氨基酸序列表明11残留proteolytically从氨基末端,留下465残留的蛋白质链NH2-terminal活跃网站半胱氨酸残基。Plasmid-encoded枯草芽孢杆菌amidophosphoribosyltransferase从大肠杆菌细胞纯化酶相比从枯草芽孢杆菌和大肠杆菌。plasmid-encoded酶相似属性amidophosphoribosyltransferase从枯草芽孢杆菌。体外酶特定的活动,免疫反应性,不稳定性,O2, Fe-S内容,NH2-terminal处理与amidophosphoribosyltransferase纯化从枯草芽孢杆菌几乎相同。 Thus E. coli correctly processed the NH2 terminus and assembled [4Fe-4S] centers in B. subtilis amidophosphoribosyltransferase although it does not perform these maturation steps on its own enzyme. Amino acid sequence comparison indicates that the B. subtilis and E. coli enzymes are homologous. Catalytic and regulatory domains were tentatively identified based on comparison with E. coli amidophosphoribosyltransferase and other phosphoribosyltransferase (Argos, P., Hanei, M., Wilson, J., and Kelley, W. (1983) J. Biol. Chem. 258, 6450-6457).