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Functional expression of His‐tagged sensory rhodopsin I in Escherichia coli
Author(s) -
Schmies Georg,
Chizhov Igor,
Engelhard Martin
Publication year - 2000
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/s0014-5793(99)01760-3
Subject(s) - halobacterium salinarum , escherichia coli , rhodopsin , bacteriorhodopsin , chemistry , biophysics , biochemistry , biology , gene , retinal , membrane
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.