Messenger RNA of opsin from bovine retina: isolation and partial sequence of the in vitro translation product.
Opsin, the apoprotein of the visual pigment rhodopsin, is synthesized on membranes of the rough endoplasmic reticulum and subsequently passes through the Golgi apparatus to the rod outer segment. This pathway parallels the early stages of biosynthesis of some secretory proteins and viral membrane glycoproteins. Most of these proteins are initially synthesized as precursor molecules with a short-lived hydrophobic extra peptide segment at the NH(2) terminus. Therefore we investigated whether or not the immediate translation product of opsin mRNA contains a similar short-lived NH(2)-terminal extra peptide. The mRNA coding for opsin was isolated from bovine retina polysomes precipitated by antibodies to opsin. The mRNA directed the cell-free synthesis of a protein comparable in size to opsin that was specifically precipitated by anti-opsin antibodies. Sequence analyses of the immunoprecipitated protein labeled with six radioactive amino acids (Met, Asn, Pro, Phe, Tyr, Val) provided the following result: [Formula: see text] (X is unknown). This partial sequence of the cell-free product corresponds exactly to the published NH(2)-terminal segment of native opsin (21 residues long) and extends beyond this region. Met-1 was shown to be the initiator methionine residue, because only the initiator [(35)S]Met-tRNA(1) (Met)-not the internal [(35)S]Met-tRNA(2) (Met)-donated the NH(2)-terminal methionine. This finding essentially rules out the possibility that Met-1 was preceded by a peptide that was rapidly cleaved. Thus opsin, and not a precursor, is the immediate product of opsin mRNA translation.
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D008970:Molecular Weight; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D012160:Retina / Q000378:metabolism*; D012168:Retinal Pigments / Q000096:biosynthesis*; D012243:Rhodopsin / Q000096:biosynthesis*
D012333:RNA, Messenger; D012168:Retinal Pigments; D012243:Rhodopsin