里氏木霉纤维二糖水解酶I和内切葡聚糖酶I核心蛋白的结晶及初步X射线研究

Crystallization and preliminary X-ray studies on the core proteins of cellobiohydrolase I and endoglucanase I from Trichoderma reesei.

作者信息

Divne C, Sinning I, Ståhlberg J, Pettersson G, Bailey M, Siika-aho M, Margolles-Clark E, Teeri T, Jones T A

机构信息

Department of Molecular Biology, Uppsala University Biomedical Centre, Sweden.

出版信息

J Mol Biol. 1993 Dec 5;234(3):905-7. doi: 10.1006/jmbi.1993.1640.

DOI:10.1006/jmbi.1993.1640

PMID:8254686

Abstract

The catalytic core domains of cellobiohydrolase I (CBHI) and endoglucanase I (EGI) from Trichoderma reesei have been crystallized using the hanging drop vapour diffusion method. In the case of CBHI, use of polyethylene glycol 20,000, and calcium chloride at low pH produced good quality single crystals suitable for X-ray studies. The crystals belong to a primitive orthorhombic space group with unit cell dimensions a = 84.0 A, b = 86.2 A, c = 111.8 A, and diffract beyond 2.0 A resolution. Bipyramidal crystals of EGI core were grown from ammonium sulphate at pH 7.5. The crystals are tetragonal, either P4(1)22 or the enantiomorph P4(3)22, with cell dimensions a = b = 101.8 A and c = 198.0 A, and at best diffract to a resolution of 2.5 A.

摘要

里氏木霉的纤维二糖水解酶I（CBHI）和内切葡聚糖酶I（EGI）的催化核心结构域已采用悬滴气相扩散法进行了结晶。对于CBHI，在低pH值下使用聚乙二醇20000和氯化钙可产生适合X射线研究的高质量单晶。这些晶体属于原始正交空间群，晶胞参数为a = 84.0 Å，b = 86.2 Å，c = 111.8 Å，并且能衍射到2.0 Å分辨率以上。EGI核心的双锥体晶体是在pH 7.5的硫酸铵中生长的。这些晶体为四方晶系，空间群为P4(1)22或对映体P4(3)22，晶胞参数a = b = 101.8 Å，c = 198.0 Å，最佳衍射分辨率为2.5 Å。