Laser-improved protein crystallization screening.
Laser-improved protein crystallization screening.
复制标题
激光改进的蛋白质结晶筛选。
DOI：
10.1107/s1744309110023857
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Yennawar,Hemant
中科院分区：
文献类型：
--
作者：
Yennawar,Neela;Denev,Sava;Gopalan,Venkataraman;Yennawar,Hemant
用核糖核酸酶B、葡萄糖脱氢酶、溶菌酶、山梨醇脱氢酶、果糖脱氢酶和肌红蛋白等6种蛋白质研究了激光照射下蛋白质结晶的筛选。用皮秒脉冲和6 mW功率的532 nm绿色圆偏振激光照射新设置的蛋白质液滴30 s，与相同条件下但没有激光照射的对照液滴相比，显示出适合晶体生长的筛选条件的数量显著改善。 对于葡萄糖脱氢酶和山梨醇脱氢酶，形成了更大和更好质量的晶体，并且提高了X射线衍射的分辨率。在核糖核酸酶B、溶菌酶和山梨醇脱氢酶的情况下，结晶速度增加。在激光照射过程中，筛选液滴中的沉淀量增加，表明蛋白质溶解度短暂降低。在优化的激光设置下，激光对晶体生长或蛋白质没有有害影响。在核糖核酸酶B和溶菌酶的情况下，晶体包装没有改变，由于激光曝光。
Screening of proteins for crystallization under laser irradiation was investigated using six proteins: ribonuclease B, glucose dehydrogenase, lysozyme, sorbitol dehydrogenase, fructose dehydrogenase and myoglobin. Shining 532 nm green circularly polarized laser light with a picosecond pulse and 6 mW power for 30 s on newly set-up protein drops showed a marked improvement in the number of screen conditions amenable for crystal growth compared with control drops under identical conditions but without laser exposure. For glucose dehydrogenase and sorbitol dehydrogenase, larger and better quality crystals were formed and the resolution of X-ray diffraction was improved. The speed of crystallization increased in the case of ribonuclease B, lysozyme and sorbitol dehydrogenase. During laser irradiation, the amount of precipitation in the screened drops increased, indicating a transient decrease in protein solubility. At the optimized laser settings, there was no deleterious effect of the laser on crystal growth or on the protein. In the cases of ribonuclease B and lysozyme the crystal packing did not change owing to the laser exposure.