Crystallization of Microbial Rhodopsins

Methods Mol Biol. 2022:2501:125-146. doi: 10.1007/978-1-0716-2329-9_6.

Abstract

Microbial rhodopsins are light-sensitive transmembrane proteins, evolutionary adapted by various organisms like archaea, bacteria, simple eukaryote, and viruses to utilize solar energy for their survival. A complete understanding of functional mechanisms of these proteins is not possible without the knowledge of their high-resolution structures, which can be primarily obtained by X-ray crystallography. This technique, however, requires high-quality crystals, growing of which is a great challenge especially in case of membrane proteins. In this chapter, we summarize methods applied for crystallization of microbial rhodopsins with the emphasis on crystallization in lipidic mesophases, also known as in meso approach. In particular, we describe in detail the methods of crystallization using lipidic cubic phase to grow both large crystals optimized for traditional crystallographic data collection and microcrystals for serial crystallography.

Keywords: Bicelles; Crystallization; In meso; Lipidic cubic phase; Membrane fusion; Microbial rhodopsins; Serial crystallography; X-ray diffraction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization / methods
  • Crystallography, X-Ray
  • Lipids / chemistry
  • Membrane Proteins* / chemistry
  • Rhodopsins, Microbial*

Substances

  • Lipids
  • Membrane Proteins
  • Rhodopsins, Microbial