title: Functional characterization of sensory rhodopsin II from Halobacterium salinarum expressed in Escherichia coli.
abstract: Sensory rhodopsin II (SRII) from Halobacterium salinarum is heterologously expressed in Escherichia coli with a yield of 3-4 mg of purified SRII per liter cell culture. UV/Vis absorption spectroscopy display bands characteristic for native SRII. The resonance Raman spectrum provides evidence for a strongly hydrogen-bonded Schiff base like in mammalian rhodopsin but unlike to the homologous pSRII from Natronobacterium pharaonis. Laser flash spectroscopy indicates that SRII in detergent as well as after reconstitution into polar lipids shows its typical photochemical properties with prolonged photocycle kinetics. The first functional heterologous expression of SRII from H. salinarum provides the basis for studies with its cognate transducer HtrII to investigate the molecular processes involved in phototransduction as well as in chemotransduction.
authors.affiliation: Research Centre Jülich, IBI-2: Structural Biology, 52425 Jülich, Germany.
authors.forename: O S
authors.initials: OS
authors.lastname: Mironova
authors.forename: R G
authors.initials: RG
authors.lastname: Efremov
authors.forename: B
authors.initials: B
authors.lastname: Person
authors.forename: J
authors.initials: J
authors.lastname: Heberle
authors.forename: I L
authors.initials: IL
authors.lastname: Budyak
authors.forename: G
authors.initials: G
authors.lastname: Büldt
authors.forename: R
authors.initials: R
authors.lastname: Schlesinger
mesh_terms: D004586:Electrophoresis; D004926:Escherichia coli / Q000235:genetics*; D019617:Halobacterium salinarum / Q000235:genetics*; D007700:Kinetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D025603:Sensory Rhodopsins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D013059:Spectrum Analysis, Raman
chemical_list: D011994:Recombinant Proteins; D025603:Sensory Rhodopsins