title: An unconventional anaerobic membrane protein production system based on Wolinella succinogenes.
abstract: In cases where membrane protein production attempts in more conventional Escherichia coli-based systems have failed, a solution is to resort to a system based on the nonpathogenic epsilon-proteobacterium Wolinella succinogenes. This approach has been demonstrated to be successful for structural and mechanistic analyses not only for homologous production of W. succinogenes membrane proteins but also for the heterologous production of membrane protein complexes from the human pathogens Helicobacter pylori and Campylobacter jejuni. The procedure to establish a system for the production of native and variant enzymes in W. succinogenes is presented in detail for the examples of the quinol:fumarate reductase and the SdhABE complexes of W. succinogenes. Subsequently, further projects using W. succinogenes as expression host are covered.
authors: Department of Structural Biology, Institute of Biophysics and Center of Human and Molecular Biology (ZHMB), Saarland University, Homburg, Germany.
authors: Michael
authors: M
authors: Lafontaine
authors: Department of Structural Biology, Institute of Biophysics and Center of Human and Molecular Biology (ZHMB), Saarland University, Homburg, Germany. Electronic address: roy.lancaster@structural-biology.eu.
authors: C Roy D
authors: CR
authors: Lancaster
mesh_terms: D002850:Chromatography, Gel; D003001:Cloning, Molecular / Q000379:methods*; D018360:Crystallography, X-Ray; D005822:Genetic Vectors / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014170:Transformation, Genetic; D016968:Wolinella / Q000235:genetics* / Q000254:growth & development
chemical_list: D008565:Membrane Proteins
keywords: Atomic model; Enzymic assays; Epsilon-proteobacteria; Heterologous expression; Membrane protein complex; Membrane protein crystallization; Membrane protein production; Pathogenic bacteria; Proteoliposomes; X-ray crystallography