title: Crystallization of membrane proteins by vapor diffusion.
abstract: X-ray crystallography remains the most robust method to determine protein structure at the atomic level. However, the bottlenecks of protein expression and purification often discourage further study. In this chapter, we address the most common problems encountered at these stages. Based on our experiences in expressing and purifying antimicrobial efflux proteins, we explain how a pure and homogenous protein sample can be successfully crystallized by the vapor diffusion method. We present our current protocols and methodologies for this technique. Case studies show step-by-step how we have overcome problems related to expression and diffraction, eventually producing high-quality membrane protein crystals for structural determinations. It is our hope that a rational approach can be made of the often anecdotal process of membrane protein crystallization.
authors.affiliation: Department of Physics and Astronomy, Iowa State University, Ames, Iowa, USA.
authors.forename: Jared A
authors.initials: JA
authors.lastname: Delmar
authors.affiliation: Department of Chemistry, Iowa State University, Ames, Iowa, USA.
authors.forename: Jani Reddy
authors.initials: JR
authors.lastname: Bolla
authors.affiliation: Department of Physics and Astronomy, Iowa State University, Ames, Iowa, USA.
authors.forename: Chih-Chia
authors.initials: CC
authors.lastname: Su
authors.affiliation: Department of Physics and Astronomy, Iowa State University, Ames, Iowa, USA; Department of Chemistry, Iowa State University, Ames, Iowa, USA. Electronic address: ewyu@iastate.edu.
authors.forename: Edward W
authors.initials: EW
authors.lastname: Yu
mesh_terms: D000818:Animals; D001419:Bacteria / Q000737:chemistry / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D018360:Crystallography, X-Ray / Q000379:methods*; D003902:Detergents / Q000737:chemistry; D004058:Diffusion; D015870:Gene Expression; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008958:Models, Molecular; D014835:Volatilization
chemical_list: D001426:Bacterial Proteins; D003902:Detergents; D008565:Membrane Proteins
keywords: Antimicrobial efflux; Membrane protein crystallization; Multidrug resistance; Vapor diffusion; X-ray crystallography