title: Functional expression of His-tagged sensory rhodopsin I in Escherichia coli.
abstract: Sensory rhodopsin I (SRI) from Halobacterium salinarum was functionally expressed in Escherichia coli and subsequently purified to homogeneity using a C-terminal His-tag anchor. Yields of 3-4 mg SRI/l cell culture can be obtained. The absorption and photocycle properties of SRI were similar if not indistinguishable from those of the homologously expressed SRI. A global fit analysis of the photocycle data and the calculation of the spectra of states provided strong evidence for the existence of an N-like intermediate.
authors.affiliation: Max-Planck-Institut für Molekulare Physiologie, Otto-Hahn Strasse 11, D-44227, Dortmund, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Schmies
authors.forename: I
authors.initials: I
authors.lastname: Chizhov
authors.forename: M
authors.initials: M
authors.lastname: Engelhard
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D019617:Halobacterium salinarum / Q000235:genetics; D025602:Halorhodopsins; D006639:Histidine / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D025603:Sensory Rhodopsins; D013053:Spectrophotometry
chemical_list: D017931:DNA Primers; D025602:Halorhodopsins; D011994:Recombinant Proteins; C061497:SRI protein, Halobacterium; D025603:Sensory Rhodopsins; D006639:Histidine; D001436:Bacteriorhodopsins