Proper cotranslational insertion of visual rhodopsin into the lipid bilayer occurs in the absence of protein translocation machinery.

Abstract

The insertion of visual opsin into membranes occurred during in vitro translation of opsin mRNA in wheat germ extract in the presence of either microsomes or liposomes. The rhodopsin that integrated into both types of membranes after regeneration with 11-cis-retinal was functionally active (in contrast to the nonincorporated protein). Opsin either cotranslationally translocated into microsomes or inserted into liposomes had equal sensitivity to proteolysis and yielded the same pattern of peptides, which differed substantially from the set of peptides produced during proteolysis of opsin not incorporated into membranes. Thus visual opsin does not require protein translocation machinery for proper insertion into the lipid bilayer.