来自大肠杆菌的重组GDP-4-酮-6-脱氧-D-甘露糖差向异构酶/还原酶的初步晶体学研究。

Preliminary crystallographic investigations of recombinant GDP-4-keto-6-deoxy-D-mannose epimerase/reductase from E. coli.

作者信息

Tonetti M, Rizzi M, Vigevani P, Sturla L, Bisso A, De Flora A, Bolognesi M

机构信息

Istituto Policattedra di Chimica Biologica, Universita' di Genova, Viale Benedetto XV, 1. I-16132 Genova, Italy.

出版信息

Acta Crystallogr D Biol Crystallogr. 1998 Jul 1;54(Pt 4):684-6. doi: 10.1107/s0907444997018970.

DOI:10.1107/s0907444997018970

PMID:9761875

Abstract

The GDP-4-keto-6-deoxy-D-mannose epimerase/reductase (GM_ER) isolated from E. coli has been overexpressed as a GST-fusion protein and purified to homogeneity. The enzyme, an NADP+(H)-binding homodimer of 70 kDa, is responsible for the production of GDP-L-fucose. GM_ER shows significant structural homology to the human erythrocyte protein FX, which is involved in blood-group glycoconjugate biosynthesis, displaying 3,5 epimerase/reductase activity on GDP-4-keto-6-deoxy-D-mannose. GM_ER has been crystallized in a trigonal crystalline form, containing one molecule per asymmetric unit, suitable for high-resolution crystallographic investigations.

摘要

从大肠杆菌中分离出的GDP-4-酮基-6-脱氧-D-甘露糖差向异构酶/还原酶（GM_ER）已作为谷胱甘肽S-转移酶（GST）融合蛋白过量表达，并纯化至均一。该酶是一种70 kDa的NADP+(H)结合同型二聚体，负责生成GDP-L-岩藻糖。GM_ER与参与血型糖缀合物生物合成的人类红细胞蛋白FX具有显著的结构同源性，对GDP-4-酮基-6-脱氧-D-甘露糖具有3,5-差向异构酶/还原酶活性。GM_ER已结晶为三方晶型，每个不对称单元包含一个分子，适合进行高分辨率晶体学研究。