Abstract
The first crystal structure of an intact potassium channel was determined in 1998. Since then, the use of X-ray crystallography for studying ion channels and fragments thereof has become an invaluable tool for elucidating the utmost basic molecular mechanisms underlying their assembly, selectivity, permeation and regulatory mechanisms. This chapter provides a contemporary overview of macromolecular X-ray crystallography. First, the principles governing protein crystal formation, the main bottleneck of crystallographic studies are explained. Then provided is an overview of bioinformatic and experimental approaches that can be employed for improving protein sample quality, thereby increasing the likelihood of obtaining crystals suitable for downstream structural studies. Next, are the procedures of the diffraction data collection experiment and structure determination. Also described are the quality metrics used for assessing the diffraction data and the structural model. Finally, as the 3D structural information is usually conveyed through 2D images, common visualization modes designed to facilitate the deduction of mechanistic insights from the determined structure are described. With the ever-growing role of structural approaches in modern ion channel studies, this chapter provides an understanding of the general workflow, capabilities and limitations of X-ray crystallography.
Original language | English |
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Title of host publication | Textbook of Ion Channels Volume I |
Subtitle of host publication | Basics and Methods |
Publisher | CRC Press |
Pages | 225-238 |
Number of pages | 14 |
Volume | 1 |
ISBN (Electronic) | 9781000857757 |
ISBN (Print) | 9780367538156 |
DOIs | |
State | Published - 1 Jan 2023 |