title: Production of human tetraspanin proteins in Escherichia coli.
abstract: Tetraspanins are found in multicellular eukaryotes and are generally thought to act as scaffolding proteins, localizing multiple proteins to a specific region of the cell membrane. Activities for tetraspanins have been identified in several fundamental processes such as motility, cell adhesion, proliferation and viral entry. Tetraspanins are also key players in cancer development and progression. However, structural and biochemical information on tetraspanins is decidely limited, due in no small part to the difficulties associated with expressing eukaryotic membrane proteins. In this study, we have used GFP fusions of a library of human tetraspanin proteins to identify growth conditions for expression in Escherichia coli. Three tetraspanin-GFP proteins could be produced at high enough levels to allow subsequent purification, paving the way for future structural and biochemical studies.
authors.affiliation: Stockholm Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, The Arrhenius Laboratories for Natural Sciences, SE-10691 Stockholm, Sweden.
authors.forename: Michael
authors.initials: M
authors.lastname: Tarry
authors.forename: Karin
authors.initials: K
authors.lastname: Skaar
authors.forename: Gunnar von
authors.initials: Gv
authors.lastname: Heijne
authors.forename: Roger R
authors.initials: RR
authors.lastname: Draheim
authors.forename: Martin
authors.initials: M
authors.lastname: Högbom
mesh_terms: D000954:Antigens, Surface / Q000096:biosynthesis* / Q000302:isolation & purification; D002021:Buffers; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli; D006801:Humans; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D012995:Solubility; D060185:Tetraspanin 24 / Q000096:biosynthesis* / Q000302:isolation & purification; D060169:Uroplakin Ib / Q000096:biosynthesis* / Q000302:isolation & purification
chemical_list: D000954:Antigens, Surface; D002021:Buffers; D003902:Detergents; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D060185:Tetraspanin 24; C086452:UPK1B protein, human; D060169:Uroplakin Ib; C074075:TM4SF1 protein, human