Crystallization, X-ray diffraction analysis and SIRAS phasing of human α-L-iduronidase
Crystallization, X-ray diffraction analysis and SIRAS phasing of human α-L-iduronidase
复制标题
人 α-L-艾杜糖醛酸酶的结晶、X 射线衍射分析和 SIRAS 定相
DOI：
10.1107/s1744309112040432
复制
发表时间：
2012
期刊：
影响因子：
--
通讯作者：
and Sakuraba H
中科院分区：
文献类型：
--
作者：
Maita N;Taniguchi H;and Sakuraba H
人溶酶体α-L-艾杜糖苷酶以酒石酸钠/酒石酸钾/聚乙二醇3350为沉淀剂进行结晶，其缺失可引起I型粘多糖病。使用同步辐射，在100 K到2.3 ä分辨率下从单晶中收集了原始数据集。晶体属于R3空间群，晶胞尺寸a=b=259.22，c=71.83 ä。为了获得相位信息，制备了汞衍生物晶体，并在汞的峰值波长采集了单波长的反常色散(SAD)数据。用单次同晶置换反常散射(SIRAS)方法成功地计算出了可解释的电子密度图。
Human lysosomal α-l-iduronidase, whose deficiency causes mucopolysaccharidosis type I, was crystallized using sodium/potassium tartrate and polyethylene glycol 3350 as a precipitant. Using synchrotron radiation, a native data set was collected from a single crystal at 100 K to 2.3 Å resolution. The crystal belonged to space group R3 with unit-cell dimensions of a = b = 259.22, c = 71.83 Å. To obtain the phase information, mercury-derivative crystals were prepared and a single-wavelength anomalous dispersion (SAD) data set was collected at the Hg peak wavelength. Phase calculation with the single isomorphous replacement with anomalous scattering (SIRAS) method successfully yielded an interpretable electron-density map.