INVESTIGATIONS ON PROTEIN CRYSTAL-GROWTH BY THE GEL ACUPUNCTURE METHOD
INVESTIGATIONS ON PROTEIN CRYSTAL-GROWTH BY THE GEL ACUPUNCTURE METHOD
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DOI：
10.1107/s0907444993014350
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发表时间：
1994-07-01
影响因子：
2.2
通讯作者：
MORENO, A
中科院分区：
文献类型：
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作者：
GARCIARUIZ, JM;MORENO, A
在这项工作中，我们探讨了凝胶针刺技术的可能性，以前提出的蛋白质单晶的生长[加西亚-鲁伊斯，莫雷诺，维埃德马和科尔（1993年）。Mater. Res. Bull.28，541-546]。该技术的主要优点是，晶体是在X射线毛细管内获得的，与经典的微扩散技术不同，它涉及一个非常简单和准确的技术安排，允许在其生长环境中连续监测晶体。特别地，我们描述了溶菌酶、伴刀豆球蛋白A和核糖核酸酶A的单晶生长。不同的起始条件已被用于在几种蛋白质和沉淀剂浓度下将这些蛋白质的单晶生长到不同类型的毛细管中。结果表明，该技术适用于蛋白质晶体生长中常用的各种沉淀剂，如大聚合物（PEG 4000和PEG 6000），有机溶剂（从甲醇到丁醇）和盐[NaCl，（NH 4）2SO 4]。还研究了该技术正确工作的毛细管内径范围。生长过程和可能的晶体运动，其次是视频显微镜。溶菌酶晶体可达3.1 mm，但平均最大线性晶体尺寸分别为2.0 mm的溶菌酶，0.4 mm的伴刀豆球蛋白A和1.2 mm的核糖核酸酶，分别。从实验设置开始测量，达到这样的尺寸的等待时间分别为72小时、10天和5天。凝胶的四甲氧基硅烷，四乙氧基硅烷，硅酸钠，琼脂，高强度琼脂和gel-gro已被测试有关其机械性能和它们与反应物的化学相互作用。最后，我们简要讨论了凝胶针刺技术的优点和合理的应用以外的晶体生长。
In this work we explore the possibilities of the gel-acupuncture technique, proposed previously for the growth of protein single crystals [Garcia-Ruiz, Moreno, Viedma & Coll (1993). Mater. Res. Bull. 28, 541-546]. The main advantage of the technique is that the crystals are obtained inside an X-ray capillary and, unlike classical microdiffusion techniques, it involves a very simple and accurate technical arrangement that permits the continuous monitoring of the crystals in their growth environment. In particular, we describe the growth of single crystals of lysozyme, concanavalin A and ribonuclease A. Different starting conditions have been used to grow single crystals of these proteins into different types of capillaries at several protein and precipitating-agent concentrations. It is demonstrated that the technique works for a wide range of precipitating agents commonly used in protein crystal growth, such as large polymers (PEG 4000 and PEG 6000), organic solvents (from methanol to butanol) and salts [NaCl, (NH4)2SO4]. The range of inner diameter of the capillaries for which the technique works correctly has been also studied. The growth process and possible crystal movement was followed by video microscopy. Lysozyme crystals up to 3.1 mm were obtained but the average maximum linear crystal sizes were 2.0 mm for lysozyme, 0.4 mm for concanavalin A and 1.2 mm for ribonuclease, respectively. The waiting times to reach such a size, measured from the set-up of the experiments, were 72 h, 10 d and 5 d, respectively. Gels of tetramethoxysilane, tetraethoxysilane, sodium silicate, agar, high-strength agar and gel-gro have been tested in relation to their mechanical properties and their chemical interaction with the reactants. Finally, we discuss briefly the advantages of the gel-acupuncture technique and plausible applications other than crystal growth.