大鼠PSD - 95蛋白鸟苷酸激酶样结构域的结晶及初步X射线衍射研究

Crystallization and preliminary X-ray diffraction studies of the guanylate kinase-like domain of PSD-95 protein from rat.

作者信息

Kim J J, Rho S H, Im Y J, Kim E J, Eom S H

机构信息

Department of Life Science, Kwangju Institute of Science and Technology, Kwangju 500-712, South Korea.

出版信息

Acta Crystallogr D Biol Crystallogr. 2001 Apr;57(Pt 4):616-7. doi: 10.1107/s0907444901001937.

DOI:10.1107/s0907444901001937

PMID:11264600

Abstract

The PSD-95 (postsynaptic density-95) protein, one of the members of the MAGUK (membrane-associated guanylate kinase) family, is composed of three PDZ domains, one SH3 domain and one guanylate kinase-like (GK) domain. The GK domain mediates the scaffolding function of PSD-95 by protein--protein interaction. Here, the GK domain was subcloned, expressed as an intein fusion protein, purified without the intein and then crystallized at room temperature by the hanging-drop vapour-diffusion method using PEG 8000 as a precipitant. The complete native data set was collected to a resolution of 2.35 A using flash-cooling. The crystals belong to the primitive tetragonal space group P4(3) (or P4(1)), with unit-cell parameters a = b = 70.03 (4), c = 37.64 (1) A.

摘要

PSD-95（突触后致密蛋白95）是MAGUK（膜相关鸟苷酸激酶）家族成员之一，由三个PDZ结构域、一个SH3结构域和一个类鸟苷酸激酶（GK）结构域组成。GK结构域通过蛋白质-蛋白质相互作用介导PSD-95的支架功能。在此，GK结构域被亚克隆，表达为内含肽融合蛋白，去除内含肽后进行纯化，然后在室温下使用PEG 8000作为沉淀剂通过悬滴气相扩散法进行结晶。使用快速冷却将完整的天然数据集收集至2.35 Å的分辨率。晶体属于原始四方空间群P4(3)（或P4(1)），晶胞参数a = b = 70.03 (4)，c = 37.64 (1) Å。