Hamana H, Moriyama H, Shinozawa T, Tanaka N
Department of Biological and Chemical Engineering, Faculty of Engineering, Gunma University, Kiryu Gunma 376, Japan.
Acta Crystallogr D Biol Crystallogr. 1999 Jan;55(Pt 1):345-6. doi: 10.1107/S0907444998008130. Epub 1999 Jan 1.
Crystallization of orotate phosphoribosyltransferase from a thermophilic organism, Thermus thermophilus, was achieved using the hanging-drop vapour-diffusion method coupled with a macroseeding starter. Small needle-like microcrystals were grown in a fresh protein solution in the presence of 2-methyl-2,4-pentanediol at 298 K or below. Although these normal temperature conditions caused stacking crystallization, an increase of temperature to 310 K permitted crystal growth. This was because of increased enzyme solubility at the higher temperature. The crystal was found to belong to the monoclinic space group P21with unit-cell parameters a = 44.4, b = 59.6, c = 67.8 A and beta = 98.3 degrees.
利用悬滴气相扩散法结合大晶种起始物,实现了嗜热栖热菌(Thermus thermophilus)中乳清酸磷酸核糖基转移酶的结晶。在298 K或更低温度下,于含有2-甲基-2,4-戊二醇的新鲜蛋白质溶液中生长出小的针状微晶。尽管这些常温条件导致了堆积结晶,但将温度升至310 K可使晶体生长。这是因为在较高温度下酶的溶解度增加。发现该晶体属于单斜空间群P21,晶胞参数为a = 44.4、b = 59.6、c = 67.8 Å,β = 98.3°。