Auerbach G, Jacob U, Grättinger M, Schurig H, Jaenicke R
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Biol Chem. 1997 Mar-Apr;378(3-4):327-9.
Phosphoglycerate kinase from the hyperthermophilic bacterium Thermotoga maritima has been co-crystallized with its substrate 3-phosphoglycerate and the ATP analogue AMP-PNP using the vapour diffusion method. Crystals were obtained from a solution containing polyethylene glycol (MW 3000/8000) as precipitating agent. A complete diffraction data set from orthorhombic crystals was collected up to 2.0 A resolution. The TmPGK crystallizes in the space group P2(1)2(1)2 (cell dimensions: a = 62.0 A, b = 76.9 A, c = 87.5 A) with one molecule in the asymmetric unit. The structure was solved by Patterson search methods using Bacillus stearothermophilus PGK as a search model and was refined to a crystallographic R factor of 22.0%. Compared to the enzyme from B. stearothermophilus, horse, pig and yeast, the Thermotoga enzyme exhibits a drastically reduced interdomain angle, similar to the one reported for PGK from Trypanosoma brucei. Here we present crystallographic data of the first high-resolution structure of a PGK in largely closed conformation, complexed with the two products of the catalyzed reaction, and, at the same time, the first PGK structure from a hyperthermophilic organism.
嗜热栖热菌（Thermotoga maritima）的磷酸甘油酸激酶已通过气相扩散法与底物3-磷酸甘油酸及ATP类似物AMP-PNP共结晶。晶体是从含有聚乙二醇（分子量3000/8000）作为沉淀剂的溶液中获得的。收集了来自正交晶体的完整衍射数据集，分辨率达到2.0 Å。嗜热栖热菌磷酸甘油酸激酶（TmPGK）结晶于空间群P2(1)2(1)2（晶胞参数：a = 62.0 Å，b = 76.9 Å，c = 87.5 Å），不对称单元中有一个分子。结构通过以嗜热脂肪芽孢杆菌（Bacillus stearothermophilus）的磷酸甘油酸激酶为搜索模型的帕特森搜索方法解析，并精修至晶体学R因子为22.0%。与嗜热脂肪芽孢杆菌、马、猪和酵母的酶相比，嗜热栖热菌的酶表现出显著减小的结构域间夹角，类似于布氏锥虫（Trypanosoma brucei）的磷酸甘油酸激酶所报道的夹角。在此，我们展示了处于大致闭合构象的磷酸甘油酸激酶的首个高分辨率晶体结构数据，该结构与催化反应的两种产物复合，同时也是来自嗜热生物的首个磷酸甘油酸激酶结构。
