在室温下于两天内生长用于X射线衍射研究的钙调蛋白晶体。

Growing calmodulin crystals for X-ray diffraction studies at room temperature in 2 days.

作者信息

Barford A M, Gilliland G L, Morgan D W

出版信息

Anal Biochem. 1986 Nov 1;158(2):361-4. doi: 10.1016/0003-2697(86)90561-0.

Abstract

Crystals of bovine brain calmodulin have been grown using a novel procedure which utilizes a mixture of alcohols as a precipitant. Crystals were grown by vapor diffusion at room temperature in the presence of 15% ethanol, 25% 2-methyl-2,4-pentanediol, 5.0 mM calcium chloride, and 10.0 mM sodium acetate buffer at pH 4.0. Crystals were visible within 4 h and grew to 0.75 X 0.3 X 0.125 mm in 2 days, without seeding. The crystals of calmodulin are isomorphous with those found by W. J. Cook, J. R. Dedman, A. R. Means, and C. E. Bugg (1980, J. Biol. Chem. 255, 8152-8153). The protein crystallizes in space group P1 with unit cell dimensions a = 29.92 A, b = 55.96 A, c = 24.75 A, alpha = 93.81 degrees, beta = 99.24 degrees, and gamma = 88.40 degrees.

摘要

利用一种使用醇类混合物作为沉淀剂的新方法培养出了牛脑钙调蛋白晶体。晶体通过在室温下的气相扩散法培养，培养体系含有15%乙醇、25% 2-甲基-2,4-戊二醇、5.0 mM氯化钙和pH为4.0的10.0 mM乙酸钠缓冲液。晶体在4小时内可见，且在未接种的情况下于2天内生长至0.75×0.3×0.125毫米。钙调蛋白晶体与W. J. 库克、J. R. 戴德曼、A. R. 米恩斯和C. E. 巴格（1980年，《生物化学杂志》255卷，8152 - 8153页）所发现的晶体同晶型。该蛋白质在空间群P1中结晶，晶胞参数为a = 29.92 Å，b = 55.96 Å，c = 24.75 Å，α = 93.81°，β = 99.24°，γ = 88.40°。