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Functional characterization of sensory rhodopsin II from Halobacterium salinarum expressed in Escherichia coli
Author(s) -
Mironova O.S.,
Efremov R.G.,
Person B.,
Heberle J.,
Budyak I.L.,
Büldt G.,
Schlesinger R.
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.05.010
Subject(s) - halobacterium salinarum , rhodopsin , escherichia coli , chemistry , biochemistry , halobacteriaceae , halorhodopsin , resonance raman spectroscopy , biology , biophysics , raman spectroscopy , bacteriorhodopsin , membrane , retinal , physics , optics , gene
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.