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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Combinatorial crystallization of an RNA-protein complex.
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Combinatorial crystallization of an RNA-protein complex.

机译：组合一个rna蛋白质的结晶复杂。

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摘要

One of the most difficult steps in X-ray crystallography of a ribonucleoprotein (RNP) complex is obtaining crystals that diffract to high resolution. This paper describes a procedure for identifying the optimal lengths of the nucleic acid components that provide high-quality crystals of the RNP. Both strands of an RNA duplex were varied in a systematic manner to generate a large number of unique RNPs that were screened for crystallization behavior. As observed in the crystallization of other nucleic acids and their complexes, the exact length of the RNA chains was found to be critical in obtaining diffraction-quality crystals, even though the relative molecular weights of the protein and RNA components were approximately 50 and approximately 10 kDa, respectively. In particular, the helix-loop-helix structure in the mRNA for the Saccharomyces cerevisiae ribosomal protein L30, which functions as an autoregulatory element for L30 expression, was synthesized as two separate RNA chains of variable length (12-14 and 15-17 nucletides). Duplex formation of these RNAs formed the asymmetric, internal loop-binding site for L30. 16 such RNA duplexes, varying by +/-1 residue at the 5' or 3' end of either chain, were used to prepare 16 unique complexes with a maltose-binding protein-L30 fusion protein. The complexes were screened against 48 standard crystallization conditions in 2304 experiments, yielding 30 conditions with single crystals in the initial screen. The most promising of these is being used for structure determination.
机译：在x射线最困难的步骤之一结晶学的核糖核蛋白(RNP)复杂的是获得晶体衍射高分辨率。为确定最优长度的提供高质量的核酸组件RNP的晶体。双系统的方式是多样的产生大量的独特rnp筛查结晶行为。观察到的其他核酸的结晶酸及其复合物,确切的长度RNA链被发现是至关重要的获得diffraction-quality晶体尽管的相对分子量蛋白质和RNA组件大约50大约10 kDa,分别。具体来说,helix-loop-helix结构酿酒酵母核糖体的信使rna蛋白质翻译,作为一种自动调整的功能翻译表达元素,合成两个独立的RNA链可变长度(12 - 14和nucletides 15 - 17日)。rna形成了不对称,内部loop-binding网站翻译。+ / - 1 5”或3 '末端的残渣要么链,被用来准备16独特的复合物的maltose-binding protein-L30融合蛋白。复合物对48筛选标准2304年结晶条件的实验中,产生30条件单一晶体初始屏幕。是用于结构的决心。

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