同步辐射在D-甘油醛-3-磷酸脱氢酶及其系列衍生物高分辨率衍射数据收集中的应用

用气相扩散法培养出 P.Versicolor龙虾肌HOLO-D-甘油醛-3-磷酸脱氢酶(HOLO一GAPDH)、149位巯基羧甲基化的衍生物(CM—GAPDH)和紫外光激发生成的新NAD荧光衍生物(IRR—GAPDH)的晶体.用同步辐射强x光光源一磷光贮屏一Weissenberg照相机系统,选择a+b或a-b晶轴为旋转轴;收集了酶及其衍生物1.8A分辨率的三套衍射数据;用WEIS程序进行了数据处理.三套数据的Rmerge分别为4.93%、6.22%和5.77%.用CAD程序对三套数据进行了比较分析.

THE APPLICATION OF SYNCHROTRON RAIATION IN DIFFRACT ON DATA COLLECTION OF D-GLYCERALDEHYDE-3-PHOSPHAIE DEHVDROGENASE AND IT'S DERIVATIVES

Helo - D - glyceraldehyde - 3 - phosphate dehyregenase HOLO - GAPDUwas purified frorr P. versicolor lobster muscle. It's active site Cys-149 carboxymethy and den-vative CM-GAPDH) and the fluorescent NAD derivative IRR-GAPDH were prepared. Using I (NH4) 2SO4 as precipitating agent, the three forms of the en me were crystallized by vapour diffusion method. The diffraction data of these three forms of the enzyme were collected with synchrotron radiation-Fuji imaging plate -weisse berg cam-era system at BL-6A2 of Photon Factory at 1.8A resolution. Rmerge factor of HOLO-GAPDH, CM-GAPDH and IRR-GAPDH are 4.93%, 6.22% and 5.77% respectively. The diffraction intensity difference between the native enzyme and it's derivatives and its structural implication arc discussed.