结核分枝杆菌H37Rv磺基转移酶的过量表达、纯化及初步X射线衍射分析
Overproduction, purification and preliminary X-ray diffraction analysis of a sulfotransferase from Mycobacterium tuberculosis H37Rv.
作者信息
Tanaka Shotaro, Moriizumi Yuuji, Kimura Makoto, Kakuta Yoshimitsu
机构信息
Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, Hakozaki 6-10-1, Fukuoka 812-8581, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Jan 1;61(Pt 1):33-5. doi: 10.1107/S1744309104022328. Epub 2004 Sep 25.
Sulfotransferase STF1 from the Mycobacterium tuberculosis H37Rv genome was overproduced in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystals diffract to 1.5 A resolution using synchrotron radiation at SPring-8. The crystals are monoclinic and belong to space group P2(1), with unit-cell parameters a = 40.86, b = 95.76, c = 48.04 A, beta = 106.43 degrees. The calculated Matthews coefficient is approximately 2.1 A3 Da(-1) assuming the presence of one molecule of STF1 in the asymmetric unit. A substrate-binding assay using a PAP-agarose column suggests that STF1 exhibits sulfotransferase activity.
结核分枝杆菌H37Rv基因组中的磺基转移酶STF1在大肠杆菌中过量表达，经纯化后采用悬滴气相扩散法进行结晶。利用SPring-8的同步辐射，晶体衍射分辨率达到1.5 Å。晶体为单斜晶系，属于空间群P2(1)，晶胞参数a = 40.86、b = 95.76、c = 48.04 Å，β = 106.43°。假设不对称单元中存在一个STF1分子，计算得到的马修斯系数约为2.1 Å3 Da-1。使用PAP-琼脂糖柱进行的底物结合试验表明，STF1具有磺基转移酶活性。
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