Structure of lobster apo-D-glyceraldehyde-3-phosphate dehydrogenase at 3.0 A resolution.
Structure of lobster apo-D-glyceraldehyde-3-phosphate dehydrogenase at 3.0 A resolution.
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3.0 A 分辨率下龙虾 apo-D-甘油醛-3-磷酸脱氢酶的结构。
DOI：
10.1016/0022-2836(80)90069-8
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发表时间：
1980
影响因子：
5.6
通讯作者：
Rossmann,MG
中科院分区：
文献类型：
--
作者：
Murthy,MR;Garavito,RM;Johnson,JE;Rossmann,MG
龙虾脱辅基甘油醛-3-磷酸脱氢酶的制备是通过先将pH降低到4.8，从而降低NAD结合能，然后在Sephadex柱上分离酶和辅酶。三斜晶体从pH 6.2的硫酸铵溶液中生长。脱辅基结构最初通过与已知的全甘油醛-3-磷酸脱氢酶分子进行比较来大致确定。然后用222分子对称性和分子置换技术对前者进行了改进。载脂蛋白和全甘油醛-3-磷酸脱氢酶之间只有微小的构象差异。Trp 193在“S环”和腺嘌呤结合口袋显示出最显着的变化。
Lobster apo-glyceraldehyde-3-phosphate dehydrogenase was prepared by first lowering the pH to 4.8, thus reducing the NAD binding energy, and then separating the enzyme and coenzyme on a Sephadex column. Triclinic crystals were grown from an ammonium sulfate solution at pH 6.2. The apo-structure was initially determined approximately by comparison with the known hologlyceraldehyde-3-phosphate dehydrogenase molecule. The former was then refined using the 222 molecular symmetry with the molecular replacement technique. Only minor conformational differences were observed between apo and holo-glyceraldehyde-3-phosphate dehydrogenase. Trp193 in the “S loop” and the adenine-binding pocket showed the most significant changes.