Tyagi Swati, Lemke Edward A
Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Curr Opin Struct Biol. 2015 Jun;32:66-73. doi: 10.1016/j.sbi.2015.02.009. Epub 2015 Mar 8.
Contemporary structural biology research promises more than just static snap-shots of molecular machineries. This goal is not just facilitated by combining different structural biology techniques, but also by new tools from the field of protein and genetic engineering, as well as from chemistry. Genetic encoding of noncanonical amino acids (ncAAs) through codon-suppression technology provides an excellent opportunity to probe biomolecules using different structural biology methods. In this article, we review the applications of ncAA incorporation into proteins for determining structural information through various techniques with the main focus on crosslinking mass spectrometry and single-molecule FRET-based techniques. Furthermore, advances and limitations of the incorporation of multiple ncAAs are discussed, with respect to design of an ideal host organism for modern and integrative structural biology research.
当代结构生物学研究带来的远不止分子机器的静态快照。实现这一目标不仅得益于不同结构生物学技术的结合，还得益于蛋白质与基因工程领域以及化学领域的新工具。通过密码子抑制技术对非标准氨基酸（ncAAs）进行遗传编码，为利用不同结构生物学方法探测生物分子提供了绝佳机会。在本文中，我们综述了将ncAAs掺入蛋白质中以通过各种技术确定结构信息的应用，主要侧重于交联质谱和基于单分子荧光共振能量转移的技术。此外，还讨论了掺入多个ncAAs在为现代综合结构生物学研究设计理想宿主生物方面的进展和局限性。
