Two-Dimensional Crystallization Procedure, from Protein Expression to Sample Preparation

• Published in: BioMed research international Vol. 2015; no. 2015; pp. 1 - 10
• Main Authors: Kuang, Qie, Hebert, Hans, Purhonen, Pasi
• Format: Journal Article
• Language: English
• Published: Cairo, Egypt Hindawi Publishing Corporation 01-01-2015; John Wiley & Sons, Inc; Hindawi Limited
• Subjects: Comparative analysis; Crystallization; Crystallization - methods; Crystallography; E coli; Lipids; Medicin och hälsovetenskap; Membrane proteins; Membrane Proteins - analysis; Membrane Proteins - chemistry; Membranes; Methods; Microscopy; Microscopy, Electron - methods; Physiological aspects; Plasmids; Proteins; Quality; Recombinant Proteins - analysis; Recombinant Proteins - chemistry; Review; Structure
• ISSN: 2314-6133; 2314-6141; 2314-6141
• DOI: 10.1155/2015/693869

Membrane proteins play important roles for living cells. Structural studies of membrane proteins provide deeper understanding of their mechanisms and further aid in drug design. As compared to other methods, electron microscopy is uniquely suitable for analysis of a broad range of specimens, from small proteins to large complexes. Of various electron microscopic methods, electron crystallography is particularly well-suited to study membrane proteins which are reconstituted into two-dimensional crystals in lipid environments. In this review, we discuss the steps and parameters for obtaining large and well-ordered two-dimensional crystals. A general description of the principle in each step is provided since this information can also be applied to other biochemical and biophysical methods. The examples are taken from our own studies and published results with related proteins. Our purpose is to give readers a more general idea of electron crystallography and to share our experiences in obtaining suitable crystals for data collection.