title: Overproduction of PIB-Type ATPases.
abstract: Understanding of the functions and mechanisms of fundamental processes in the cell requires structural information. Structural studies of membrane proteins typically necessitate large amounts of purified and preferably homogenous target protein. Here, we describe a rapid overproduction and purification strategy of a bacterial PIB-type ATPase for isolation of milligrams of target protein per liter Escherichia coli cell culture, with a final quality of the sample which is sufficient for generating high-resolution crystals.
authors.affiliation: School of Medicine, Tsinghua University, Beijing, China.
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Aarhus C, Denmark.
authors.forename: Oleg
authors.initials: O
authors.lastname: Sitsel
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200, Copenhagen, Denmark.
authors.forename: Kaituo
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200, Copenhagen, Denmark. pontus@sund.ku.dk.|Department of Experimental Medical Science, Lund University, Sölvegatan 19, SE-22184, Lund, Sweden. pontus@sund.ku.dk.
authors.forename: Pontus
authors.initials: P
authors.lastname: Gourdon
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology*; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D008967:Molecular Biology / Q000379:methods*
chemical_list: D008565:Membrane Proteins; D000251:Adenosine Triphosphatases
keywords: Membrane preparation; Membrane proteins; Nickel affinity chromatography; Overproduction; P-type ATPase; Size-exclusion chromatography