Holcomb Joshua, Spellmon Nicholas, Zhang Yingxue, Doughan Maysaa, Li Chunying, Yang Zhe
Department of Microbiology, Immunology, and Biochemistry, Wayne State University School of Medicine, Detroit, MI, USA.
Center for Molecular and Translational Medicine, Georgia State University, Atlanta, GA, USA.
AIMS Biophys. 2017;4(4):557-575. doi: 10.3934/biophy.2017.4.557. Epub 2017 Sep 26.
To date, X-ray crystallography remains the gold standard for the determination of macromolecular structure and protein substrate interactions. However, the unpredictability of obtaining a protein crystal remains the limiting factor and continues to be the bottleneck in determining protein structures. A vast amount of research has been conducted in order to circumvent this issue with limited success. No single method has proven to guarantee the crystallization of all proteins. However, techniques using antibody fragments, lipids, carrier proteins, and even mutagenesis of crystal contacts have been implemented to increase the odds of obtaining a crystal with adequate diffraction. In addition, we review a new technique using the scaffolding ability of PDZ domains to facilitate nucleation and crystal lattice formation. Although in its infancy, such technology may be a valuable asset and another method in the crystallography toolbox to further the chances of crystallizing problematic proteins.
迄今为止，X射线晶体学仍然是确定大分子结构和蛋白质底物相互作用的金标准。然而，获得蛋白质晶体的不可预测性仍然是限制因素，并且仍然是确定蛋白质结构的瓶颈。为了规避这个问题，已经进行了大量研究，但取得的成功有限。没有一种单一方法被证明能保证所有蛋白质都结晶。然而，已经采用了使用抗体片段、脂质、载体蛋白甚至晶体接触诱变等技术来增加获得具有足够衍射能力晶体的几率。此外，我们还综述了一种利用PDZ结构域的支架能力来促进成核和晶格形成的新技术。尽管这项技术尚处于起步阶段，但它可能是一项有价值的资产，也是晶体学工具箱中的另一种方法，可进一步提高使有问题的蛋白质结晶的机会。
