Purification of histidine tagged bacteriorhodopsin, pharaonis halorhodopsin and pharaonis sensory rhodopsin II functionally expressed in Escherichia coli

FEBS Letters

Wegener, Ansgar A¹  Engelhard, Martin¹  Hohenfeld, Ilja P¹ 

[1] Max-Planck-Institut für molekulare Physiologie, Rheinlanddamm 201, 44139 Dortmund, Germany

关键词: Bacterial rhodopsin;    Phoborhodopsin;    Photocycle;    Eubacterial expression;    Mass spectrometry;    Immunogold labelling;    BR;    bacteriorhodopsin;    PHR;    pharaonis halorhodopsin;    PSRII;    pharaonis sensory rhodopsin II;    PSOII;    pharaonis sensory opsin II;    ESI;    electron spray ionization;    IPTG;    isopropyl-1-thio-β-galactoside;    TFE;    2;    2;    2-trifluoroethanol;   

DOI  :  10.1016/S0014-5793(98)01659-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Bacteriorhodopsin (BR) from Halobacterium salinarum as well as halorhodopsin (pHR) and sensory rhodopsin II (pSRII) from Natronobacterium pharaonis were functionally expressed in E. coli using the method of Shimono et al. [FEBS Lett. (1997) 420, 54–56]. The histidine tagged proteins were purified with yields up to 1.0 mg/l cell culture and characterized by ESI mass spectrometry and their photocycle. The pSRII and pHR photocycles were indistinguishable from the wild type proteins. The BR photocycle was considerably prolonged. pSOII is located in the cytoplasmic membrane and the C-terminus is oriented towards the cytoplasm as determined by immunogold labelling.

【 授权许可】

Unknown