罗望子Kunitz型胰蛋白酶抑制剂与猪胰蛋白酶复合物的结晶及初步X射线衍射分析
Crystallization and preliminary X-ray diffraction analysis of the complex of Kunitz-type tamarind trypsin inhibitor and porcine pancreatic trypsin.
作者信息
Tomar Sakshi, Patil Dipak N, Datta Manali, Tapas Satya, Chaudhary Anshul, Sharma Ashwani K, Tomar Shailly, Kumar Pravindra
机构信息
Department of Biotechnology, Indian Institute of Technology, Roorkee, India.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Nov 1;65(Pt 11):1179-81. doi: 10.1107/S1744309109041694. Epub 2009 Oct 30.
Abstract
The complex of Tamarindus indica Kunitz-type trypsin inhibitor and porcine trypsin has been crystallized by the sitting-drop vapour-diffusion method using ammonium acetate as precipitant and sodium acetate as buffer. The homogeneity of complex formation was checked by size-exclusion chromatography and further confirmed by reducing SDS-PAGE. The crystals diffracted to 2.0 angstrom resolution and belonged to the tetragonal space group P4(1), with unit-cell parameters a = b = 57.1, c = 120.1 angstrom. Preliminary X-ray diffraction analysis indicated the presence of one unit of inhibitor-trypsin complex per asymmetric unit, with a solvent content of 45%.
摘要
罗望子Kunitz型胰蛋白酶抑制剂与猪胰蛋白酶的复合物已通过坐滴气相扩散法结晶，使用乙酸铵作为沉淀剂，乙酸钠作为缓冲液。通过尺寸排阻色谱法检查复合物形成的均一性，并通过还原SDS-PAGE进一步确认。晶体衍射至2.0埃分辨率，属于四方晶系空间群P4(1)，晶胞参数a = b = 57.1，c = 120.1埃。初步X射线衍射分析表明每个不对称单元存在一个抑制剂-胰蛋白酶复合物单元，溶剂含量为45%。
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