title: SNARE-fusion mediated insertion of membrane proteins into native and artificial membranes.
abstract: Membrane proteins carry out functions such as nutrient uptake, ATP synthesis or transmembrane signal transduction. An increasing number of reports indicate that cellular processes are underpinned by regulated interactions between these proteins. Consequently, functional studies of these networks at a molecular level require co-reconstitution of the interacting components. Here, we report a SNARE protein-based method for incorporation of multiple membrane proteins into artificial membrane vesicles of well-defined composition, and for delivery of large water-soluble substrates into these vesicles. The approach is used for in vitro reconstruction of a fully functional bacterial respiratory chain from purified components. Furthermore, the method is used for functional incorporation of the entire F1F0 ATP synthase complex into native bacterial membranes from which this component had been genetically removed. The novel methodology offers a tool to investigate complex interaction networks between membrane-bound proteins at a molecular level, which is expected to generate functional insights into key cellular functions.
authors.affiliation: Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Gustav
authors.initials: G
authors.lastname: Nordlund
authors.affiliation: Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Peter
authors.initials: P
authors.lastname: Brzezinski
authors.affiliation: 1] Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden [2] Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
authors.forename: Christoph
authors.initials: C
authors.lastname: von Ballmoos
mesh_terms: D045304:Cytochromes c / Q000737:chemistry; D022162:Cytoplasmic Vesicles / Q000737:chemistry*; D004579:Electron Transport; D042963:Electron Transport Complex II / Q000737:chemistry; D004926:Escherichia coli; D008081:Liposomes; D008567:Membranes, Artificial; D006180:Proton-Translocating ATPases / Q000737:chemistry; D012242:Rhodobacter sphaeroides; D050600:SNARE Proteins / Q000737:chemistry*
chemical_list: D008081:Liposomes; D008567:Membranes, Artificial; D050600:SNARE Proteins; D045304:Cytochromes c; D042963:Electron Transport Complex II; D006180:Proton-Translocating ATPases