结构生物学家的分子工程工具箱。

IF 7.2 2区 生物学 Q1 BIOPHYSICS
Galia T Debelouchina, Tom W Muir
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引用次数: 42

摘要

令人兴奋的新技术发展推动了结构生物学的边界,并使生物大分子和组装的研究成为可能,这在不久之前是不可想象的。然而,结构生物学家窥探复杂分子世界的能力增强,也对蛋白质工程师在试管中重现这种复杂性的能力提出了新的要求。考虑到这一挑战,我们回顾了现代分子工程工具箱的内容,这些工具箱允许以特定位点和化学定义良好的方式操纵蛋白质。因此,我们涵盖了与半胱氨酸和其他天然氨基酸修饰相关的概念,天然化学连接,基于蛋白质和排序酶的方法,琥珀抑制以及化学和酶的生物偶联策略。我们还描述了如何使用这些工具来帮助x射线晶体学、核磁共振、低温电子显微镜和动态相互作用研究的方法学发展。我们希望这本专著能够激励结构生物学家和蛋白质工程师将这些工具应用于新的系统,并增强和扩大他们的范围,以应对理解细胞过程和疾病的分子基础方面的突出挑战。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A molecular engineering toolbox for the structural biologist.

A molecular engineering toolbox for the structural biologist.

A molecular engineering toolbox for the structural biologist.

Exciting new technological developments have pushed the boundaries of structural biology, and have enabled studies of biological macromolecules and assemblies that would have been unthinkable not long ago. Yet, the enhanced capabilities of structural biologists to pry into the complex molecular world have also placed new demands on the abilities of protein engineers to reproduce this complexity into the test tube. With this challenge in mind, we review the contents of the modern molecular engineering toolbox that allow the manipulation of proteins in a site-specific and chemically well-defined fashion. Thus, we cover concepts related to the modification of cysteines and other natural amino acids, native chemical ligation, intein and sortase-based approaches, amber suppression, as well as chemical and enzymatic bio-conjugation strategies. We also describe how these tools can be used to aid methodology development in X-ray crystallography, nuclear magnetic resonance, cryo-electron microscopy and in the studies of dynamic interactions. It is our hope that this monograph will inspire structural biologists and protein engineers alike to apply these tools to novel systems, and to enhance and broaden their scope to meet the outstanding challenges in understanding the molecular basis of cellular processes and disease.

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来源期刊
Quarterly Reviews of Biophysics
Quarterly Reviews of Biophysics 生物-生物物理
CiteScore
12.90
自引率
1.60%
发文量
16
期刊介绍: Quarterly Reviews of Biophysics covers the field of experimental and computational biophysics. Experimental biophysics span across different physics-based measurements such as optical microscopy, super-resolution imaging, electron microscopy, X-ray and neutron diffraction, spectroscopy, calorimetry, thermodynamics and their integrated uses. Computational biophysics includes theory, simulations, bioinformatics and system analysis. These biophysical methodologies are used to discover the structure, function and physiology of biological systems in varying complexities from cells, organelles, membranes, protein-nucleic acid complexes, molecular machines to molecules. The majority of reviews published are invited from authors who have made significant contributions to the field, who give critical, readable and sometimes controversial accounts of recent progress and problems in their specialty. The journal has long-standing, worldwide reputation, demonstrated by its high ranking in the ISI Science Citation Index, as a forum for general and specialized communication between biophysicists working in different areas. Thematic issues are occasionally published.
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