Schlagenhauf E, Etges R, Metcalf P
European Molecular Biology Laboratory, Heidelberg, Germany.
Proteins. 1995 May;22(1):58-66. doi: 10.1002/prot.340220109.
The membrane-bound GPI-anchored zinc metalloproteinase leishmanolysin purified from Leishmania major promastigotes has been crystallized in its mature form. Two crystal forms of leishmanolysin have been grown by the vapor diffusion method using 2-methyl-2,4-pentanediol as the precipitant. Macroseeding techniques were employed to produce large single crystals. Protein microheterogeneity in molecular size and charge was incorporated into both crystal forms. The tetragonal crystal form belongs to the space group P4(1)2(1)2 or the enantiomorph P4(3)2(1)2, has unit cell parameters of a = b = 63.6 A, c = 251.4 A, and contains one molecule per asymmetric unit. The second crystal form is monoclinic, space group C2, with unit cell dimensions a = 107.2 A, b = 90.6 A, c = 70.6 A, beta = 110.6 degrees, and also contains one molecule per asymmetric unit. Both crystal forms diffract X-rays beyond 2.6 A resolution and are suitable for X-ray analysis. Native diffraction data sets have been collected and the structure determination of leishmanolysin using a combination of the isomorphous replacement and the molecular replacement methods is in progress.
从硕大利什曼原虫前鞭毛体中纯化得到的膜结合糖基磷脂酰肌醇锚定锌金属蛋白酶利什曼溶素已结晶为成熟形式。利用2-甲基-2,4-戊二醇作为沉淀剂，通过气相扩散法培养出了利什曼溶素的两种晶体形式。采用宏观接种技术来制备大的单晶。两种晶体形式均包含了分子大小和电荷方面的蛋白质微异质性。四方晶型属于空间群P4(1)2(1)2或其对映体P4(3)2(1)2，晶胞参数为a = b = 63.6 Å，c = 251.4 Å，每个不对称单元包含一个分子。第二种晶体形式为单斜晶系，空间群为C2，晶胞尺寸a = 107.2 Å，b = 90.6 Å，c = 70.6 Å，β = 110.6°，每个不对称单元同样包含一个分子。两种晶体形式的X射线衍射分辨率均超过2.6 Å，适用于X射线分析。已收集了天然衍射数据集，目前正在结合同晶置换和分子置换方法进行利什曼溶素的结构测定。
