Expression of the archaebacterial bacterio-opsin gene with and without signal sequences in Escherichia coli: the expressed proteins are located in the membrane but bind retinal poorly.

In a further effort to obtain functional expression of the bacterio-opsin gene (bop) in Escherichia coli, the bop gene with E. coli signal sequences as well as the bop gene with the native presequence were expressed in E. coli. The location of the expressed products in the E. coli cell and their processing and folding to a structure that binds retinal as in Halobacterium halobium were investigated. All the expressed proteins were in the membrane. The proteins were largely unprocessed, and they were distributed between the outer and the inner membrane. The processed fractions, which were minor, were exclusively in the inner membrane. The processed proteins bound exogenously added all-trans-retinal but only partially, indicating that these proteins were present in at least two folded states. IMAGES: