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The effects of temperature and NaCl concentration on tetragonal lysozyme face growth rates

机译:温度和NaCl浓度对四方溶菌酶面生长速率的影响

摘要

Measurements were made of the (110) and (101) face growth rates of the tetragonal form of hen egg white lysozyme at 0.1M sodium acetate buffer, pH 4.0, from 4 to 22 C and with 3.0%, 5.0%, and 7.0% NaCl used as the precipitating salt. The data were collected at supersaturation ratios ranging from approximately 4 to approximately 63. Both decreasing temperature and increasing salt concentrations shifted plots of the growth rate versus C/C(sat) to the right, i.e. higher supersaturations were required for comparable growth rates. The observed trends in the growth data are counter to those expected from the solubility data. If tetragonal lysozyme crystal growth is by addition of ordered aggregates from the solution, then the observed growth data could be explained as a result of the effects of lowered temperature and increased salt concentration on the kinetics and equilibrium processes governing protein-protein interactions in solution. The data indicate that temperature would be a more tractable means of controlling the growth rate for tetragonal lysozyme crystals contrary to the usual practice in, e.g., vapor diffusion protein crystal growth, where both the precipitant and protein concentrations are simultaneously increased. However, the available range for control is dependent upon the protein concentration, with the greatest growth rate control being at the lower concentration.
机译:在0.1M乙酸钠缓冲液,pH 4.0、4至22℃以及3.0%,5.0%和7.0%的条件下,测量了四方形式的鸡蛋清溶菌酶的(110)和(101)面生长速率。 NaCl用作沉淀盐。以大约4到大约63的过饱和率收集数据。降低温度和增加盐浓度都使生长速率与C / C(sat)的关系图向右移动,即,可比的生长速率需要更高的过饱和度。生长数据中观察到的趋势与溶解度数据中预期的趋势相反。如果四方溶菌酶晶体的生长是通过添加溶液中有序的聚集体,则观察到的生长数据可以解释为温度降低和盐浓度增加对控制溶液中蛋白质间相互作用的动力学和平衡过程的影响。数据表明温度将是控制四方溶菌酶晶体生长速率的更容易控制的手段,这与例如蒸气扩散蛋白质晶体生长中的常规实践相反,在该实践中,沉淀剂和蛋白质浓度都同时增加。但是,可控制的范围取决于蛋白质的浓度,最大的生长速率控制在较低的浓度。

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