Crystal structure of Enterococcus hirae enolase at 2.8 Å resolution
Crystal structure of Enterococcus hirae enolase at 2.8 Å resolution
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DOI：
10.1093/jb/mvg104
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发表时间：
2003-06-01
影响因子：
2.7
通讯作者：
Shirakihara, Y
中科院分区：
文献类型：
--
作者：
Hosaka, T;Meguro, T;Shirakihara, Y
我们报道了一种烯醇化酶的晶体结构，这是革兰氏阳性细菌中首次报道的结构测定。我们分离了烯醇化酶基因并测定了其碱基序列，由DNA序列推断出该烯醇化酶由431个氨基酸组成。氨基酸序列与真核生物和原核生物的烯醇化酶非常相似，分别与大肠杆菌和酵母的烯醇化酶相同65%和50%。从hirae裂解物制备烯醇化酶得到的晶体每一不对称单元含有一个二聚体。在SPring-8同步辐射源上获得了2.8埃分辨率的x射线衍射图。晶体属于空间群14，晶胞尺寸为a = b = 153.5埃，c = 90.7埃。hirae、酵母、大肠杆菌和龙虾的烯醇化酶结构非常相似。E. hirae烯醇化酶呈“开放”构象。结构上与其他烯醇化酶最不同的区域是环L4（132-140）和L3（244-265）。考虑到这些环相对于活性位点的位置，它们似乎没有直接参与功能。我们的研究结果表明，糖酵解途径中一个重要酶的三维结构在真核生物和原核生物（包括革兰氏阳性细菌）中是进化保守的。
We report the crystal structure of an enolase from Enterococcus hirae, which is the first report of a structure determination among gram-positive bacteria. We isolated the enolase gene and determined the base sequence The amino acid sequence deduced from the DNA sequence suggests that this enolase is composed of 431 amino acids. The amino acid sequence is very similar to those of enolases from eukaryotic and prokaryotic organisms, being 65% and 50% identical to enolases from Escherichia coli and yeast, respectively. The enolase prepared from E. hirae lysate yielded crystals containing one dimer per asymmetric unit. X-ray diffraction patterns were obtained at 2.8 Angstrom resolution on a SPring-8 synchrotron radiation source. Crystals belong to space group 14 with unit cell dimensions of a = b = 153.5 Angstrom, c = 90.7 Angstrom. The E. hirae, yeast, E. coli and lobster enolase structures are very similar. The E. hirae enolase takes an "Open" conformation. The regions in the structure that differ most from other enolases are loops L4 (132-140) and L3 (244-265). Considering the positions of these loops relative to the active site, they seem to have no direct involvement in function. Our findings show that the three dimensional structure of an important enzyme in the glycolytic pathway is evolutionarily conserved among eukaryotes and prokaryotes, including gram-positive bacteria.