Davis E O, Henderson P J
Department of Biochemistry, University of Cambridge, United Kingdom.
J Biol Chem. 1987 Oct 15;262(29):13928-32.
The gene xylE, coding for xylose-proton symport in Escherichia coli, was cloned and its DNA sequence determined. The cloning strategy utilized lambda placMu insertions and exploited the proximity of xylE to malB. A 2.8-kilobase HincII fragment of cloned DNA restored [14C]xylose transport and xylose-proton symport activities to a xylose transport-negative strain. The xylE gene was identified as a 1473-base pair open reading frame, located 373 base pairs downstream of malG, encoding a hydrophobic protein of Mr 53,607. The amino acid sequence of XylE bore little resemblance to the lactose-proton LacY symporter or melibiose-sodium MelB symporter, but a high degree of homology was found with the arabinose-proton AraE symporter of E. coli and glucose transport proteins of mammals. Structural analyses and comparisons suggest that 12 membrane-spanning segments may occur in the XylE protein.
编码大肠杆菌中木糖 - 质子同向转运蛋白的基因xylE被克隆并测定了其DNA序列。克隆策略利用了λplacMu插入,并利用了xylE与malB的邻近性。克隆DNA的一个2.8千碱基的HincII片段将[14C]木糖转运和木糖 - 质子同向转运活性恢复到木糖转运阴性菌株。xylE基因被鉴定为一个1473碱基对的开放阅读框,位于malG下游373个碱基对处,编码一个Mr为53,607的疏水蛋白。XylE的氨基酸序列与乳糖 - 质子LacY同向转运蛋白或蜜二糖 - 钠MelB同向转运蛋白几乎没有相似之处,但与大肠杆菌的阿拉伯糖 - 质子AraE同向转运蛋白和哺乳动物的葡萄糖转运蛋白有高度同源性。结构分析和比较表明,XylE蛋白中可能存在12个跨膜区段。