The Role of Small Molecule Additives and Chemical Modification in Protein Crystallization
The Role of Small Molecule Additives and Chemical Modification in Protein Crystallization
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DOI：
10.1021/cg101308r
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发表时间：
2011-05-01
影响因子：
3.8
通讯作者：
Larson, S. B.
中科院分区：
文献类型：
--
作者：
McPherson, A.;Nguyen, C.;Larson, S. B.
促进蛋白质结晶的另一种方法是增强大分子之间的分子间接触或消除分子间相互作用或与可能抑制结晶的溶剂的相互作用。已经采用了位点特异性突变，以及通过遗传或蛋白水解手段进行的截断。然而，也存在重大问题。由于目标大分子的结构未知，因此可能没有良好的突变体或截短体设计基础。此外，该方法要求通过重组 DNA 技术生产蛋白质，但事实往往并非如此。我们试图通过基于两个想法的实验来解决这些问题。首先，在结晶筛选过程中，可以将多种常规小分子系统地引入母液中。通过掺入晶格，小分子提供的额外分子间相互作用可能会增强晶体成核和生长。我们正在追求的第二种方法是使用传统的蛋白质化学对各种氨基酸侧链进行化学修饰。我们相信，在某些情况下，化学修饰的蛋白质可能会被诱导结晶或比天然蛋白质更好地结晶。
An alternative approach to promoting the crystallization of proteins is to enhance intermolecular contacts between macromolecules or to eliminate intermolecular interactions or interactions with solvent that might inhibit crystallization. Site-specific mutations have been employed, as have truncations by genetic or proteolytic means. There are, however, significant problems. Because the structure of the target macromolecule is unknown, there may be no good basis for the design of mutants or truncations. In addition, the approach requires that the protein be produced by recombinant DNA technology, which is frequently not the case. We have attempted to address these issues by initiating experiments based on two ideas. The first is that a wide variety of conventional small molecules might be systematically introduced into mother liquors during crystallization screening. By incorporation into the crystal lattice, the additional intermolecular interactions that the small molecules provide might enhance crystal nucleation and growth. A second approach that we are pursuing is the chemical modification of various amino acid side chains using traditional protein chemistry. We believe that in some cases chemically modified proteins might be induced to crystallize or crystallize better than the native protein.