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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >An approach to crystallizing. proteins by synthetic symmetrization
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An approach to crystallizing. proteins by synthetic symmetrization

机译：结晶的方法。合成对称蛋白

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Previous studies of symmetry preferences in protein crystals suggest that symmetric proteins, such as homodimers, might crystallize more readily on average than asymmetric, monomeric proteins. Proteins that are naturally monomeric can be made homodimeric artificially by forming disulfide bonds between individual cysteine residues introduced by mutagenesis. Furthermore, by creating a variety of single-cysteine mutants, a series of distinct synthetic dimers can be generated for a given protein of interest, with each expected to gain advantage from its added symmetry and to exhibit a crystallization behavior distinct from the other constructs. This strategy was tested on phage T4 lysozyme, a protein whose crystallization as a monomer has been studied exhaustively. Experiments on three single-cysteine mutants, each prepared in dimeric form, yielded numerous novel crystal forms that cannot be realized by monomeric lysozyme. Six new crystal forms have been characterized. The results suggest that synthetic symmetrization may be a useful approach for enlarging the search space for crystallizing proteins.
机译：先前对蛋白质晶体中对称性偏好的研究表明，对称蛋白质（例如同二聚体）比不对称单体蛋白质平均更容易结晶。通过诱变引入的单个半胱氨酸残基之间形成二硫键，可以人工将天然单体蛋白制成同型二聚体。此外，通过创建各种单半胱氨酸突变体，可以为给定的目标蛋白质生成一系列不同的合成二聚体，每个有望从其增加的对称性中获得优势，并展现出不同于其他构建体的结晶行为。此策略已在噬菌体T4溶菌酶上进行了测试，该蛋白的单体结晶已得到详尽研究。对三个均以二聚体形式制备的单半胱氨酸突变体进行的实验产生了单体溶菌酶无法实现的许多新型晶体形式。表征了六种新的晶体形式。结果表明，合成对称化可能是扩大蛋白质结晶搜索空间的有用方法。

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