Cowan-Jacob Sandra W, Kaufmann Markus, Anselmo Anthony N, Stark Wilhelm, Grütter Markus G
Novartis Pharma AG, Core Technology Area, Analytics and Biomolecular Structure, CH-4002 Basel, Switzerland.
Acta Crystallogr D Biol Crystallogr. 2003 Dec;59(Pt 12):2218-27. doi: 10.1107/s0907444903020493. Epub 2003 Nov 27.
The crystal structure of the tetrameric form of D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) isolated from rabbit muscle was solved at 2.4 A resolution after careful dynamic light-scattering experiments to find a suitable buffer for crystallization trials. The refined model has a crystallographic R factor of 20.3%. Here, the first detailed model of a mammalian GAPDH is presented. The cofactor NAD(+) (nicotinamide adenine dinucleotide) is bound to two subunits of the tetrameric enzyme, which is consistent with the negative cooperativity of NAD(+) binding to this enzyme. The structure of rabbit-muscle GAPDH is of interest because it shares 91% sequence identity with the human enzyme; human GAPDH is a potential target for the development of anti-apoptotic drugs. In addition, differences in the cofactor-binding pocket compared with the homology-model structure of GAPDH from the malaria parasite Plasmodium falciparum could be exploited in order to develop novel selective and potential antimalaria drugs.
在经过仔细的动态光散射实验以找到适合结晶试验的缓冲液后，从兔肌肉中分离出的四聚体形式的D -甘油醛-3-磷酸脱氢酶（GAPDH）的晶体结构以2.4埃的分辨率得到解析。精修后的模型的晶体学R因子为20.3%。在此，展示了首个哺乳动物GAPDH的详细模型。辅因子NAD⁺（烟酰胺腺嘌呤二核苷酸）与四聚体酶的两个亚基结合，这与NAD⁺与该酶结合的负协同性一致。兔肌肉GAPDH的结构令人感兴趣，因为它与人类酶具有91%的序列同一性；人类GAPDH是抗凋亡药物开发的潜在靶点。此外，与疟原虫恶性疟原虫GAPDH的同源模型结构相比，辅因子结合口袋的差异可被利用来开发新型的选择性潜在抗疟疾药物。
