NaCl对液-液扩散法生长溶菌酶晶体的影响
Effect of NaCl on Liquid/Liquid Diffusion Lysozyme Crystal Growth
查看参考文献23篇
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文摘
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用动态光散射法研究了不同浓度NaCl对液—液扩散法生长溶菌酶晶体的影响，并测量了晶体生长前后体系的Zeta电势．结果表明，NaCl浓度较高时，在溶菌酶溶液—凝胶界面处会发生液液分层现象，溶液中一直存在较大的聚集体，生长出的晶体质量较差．而在合适的NaCl浓度下，随着溶液Zeta电势降低，溶液中溶菌酶的大的聚集体发生解聚集，生长出的晶体质量较高． |
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其他语种文摘
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Liquid/liquid diffusion is one of the often-used methods in protein crystal growth, which is affected by precipitant concentration. The knowledge of how the precipitant affects the protein crystal growth process is useful to improve the crystal quality. Dynamic light scattering is a powerful tool in studying crystal growth. It is used in this work to reveal the interaction between NaCl and lysozyme molecules/aggregates, which in some degree, can also be described by Zeta potential of protein solution. Concentrations of lysozyme are 15, 25, 35 mg'mL"1, and those of NaCl in agarose are 0.12 kg'L"1 and 0.10 kg-L"1. The results show that there exists liquid/liquid demixing at lysozyme solution-agarose gel interface when NaCl concentration is 0.12 kg'L , and some "sea urchin liked crystals appear. When NaCl concentration decreases to 0.10 kg'L"1, the "sea urchin" liked crystals disappear and the crystals are all tetragonal. From the dynamic light scattering results, It is believed that the liquid/liquid demixing at the lysozyme solution-agarose gel interface has changed the distribution of NaCl in lysozyme solution by us. So, both existence of large aggregates (larger than 1 jum) of lysozyme and the appearance of "sea urchin" liked crystals are all for lacking of efficient high concentration of NaCl. The Zeta potential of lysozyme solution under several conditions has been measured in our experiments. The Zeta potential decreases at the appropriate crystal growth condition, which reveals that more Cl" are combined with lysozyme molecules. The more combined Cl" is, the smaller the lysozyme aggregates in solution are. That accords with the results of dynamic light scattering. |
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来源
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化学学报
,2003,61(4):520-525 【核心库】
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关键词
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液-液扩散法
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溶菌酶晶体
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晶体生长
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动态光散射法
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NaCl浓度
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地址
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中国科学院力学研究所, 国家微重力重点实验室, 北京, 100080
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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0567-7351 |
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学科
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生物化学 |
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基金
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国家自然科学基金
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国家攀登计划项目
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文献收藏号
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CSCD:1152536
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