Schmidt-Krey I, Lundqvist G, Morgenstern R, Hebert H
Department of Biosciences, Karolinska Institutet, Huddinge, S-141 57, Sweden.
J Struct Biol. 1998 Oct;123(2):87-96. doi: 10.1006/jsbi.1998.4018.
Various crystallization parameters were investigated to obtain two-dimensional crystals of the detoxification enzyme microsomal glutathione transferase for structural analysis by electron crystallography. The protein was crystallized by reconstitution of the solubilized trimer into proteoliposomes. Crystallization occurs when minimal amounts of lipid in the range of three lipid molecules per protein trimer are added to the dialysate. Once crystals were obtained, the effect of several parameters on the crystallization was determined. The temperature and initial detergent concentration were found to be crucial parameters in influencing the size of the crystals, and conclusions could be drawn about the rate dependence of the crystallization process. Two highly ordered crystal forms, which are suitable for structural analysis by electron crystallography, were obtained under the two-dimensional crystallization conditions described here.
研究了各种结晶参数，以获得解毒酶微粒体谷胱甘肽转移酶的二维晶体，用于通过电子晶体学进行结构分析。通过将溶解的三聚体重构成蛋白脂质体来使蛋白质结晶。当向透析液中添加最低量的脂质（范围为每个蛋白质三聚体三个脂质分子）时会发生结晶。一旦获得晶体，就确定了几个参数对结晶的影响。发现温度和初始去污剂浓度是影响晶体大小的关键参数，并且可以得出关于结晶过程速率依赖性的结论。在此描述的二维结晶条件下获得了两种高度有序的晶体形式，它们适用于通过电子晶体学进行结构分析。
