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. 1979 Jun;76(6):2654–2658. doi: 10.1073/pnas.76.6.2654

Messenger RNA of opsin from bovine retina: Isolation and partial sequence of the in vitro translation product

Israel Schechter *, Yigal Burstein †, Ronald Zemell *,‡, Etty Ziv *, Frida Kantor *, David S Papermaster *,§
PMCID: PMC383666  PMID: 288054

Abstract

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 NH2 terminus. Therefore we investigated whether or not the immediate translation product of opsin mRNA contains a similar short-lived NH2-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 NH2-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 [35S]Met-tRNA1Met—not the internal [35S]Met-tRNA2Met—donated the NH2-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.

Keywords: rough endoplasmic reticulum, precursor protein, hydrophobic NH2-terminal extra piece, initiator methionine

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Selected References

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