Pappin, D. J., Findlay, J. B., Eliopoulos, E., Brett, M. (February 1984) Sequence variability in the retinal-attachment domain of mammalian rhodopsins. Biochem J, 217 (3). pp. 605-13. ISSN 0264-6021 (Print)0264-6021 (Linking)
Abstract
Ovine rhodopsin was regenerated with 11-cis-[15-3H]retinal and cleaved in situ by Staphylococcus aureus V8 proteinase to give two membrane-bound fragments of Mr 27 000 (V8-L) and 12 000 (V8-S). After purification of the proteolysed complex by affinity chromatography with concanavalin A-Sepharose 4B, [3H]retinal was covalently linked to the protein by reduction with borohydride. The purified [3H]-retinyl V8-S fragment was cleaved with CNBr and trifluoroacetic acid, the resulting peptides resolved by gel filtration and the [3H]retinyl peptide sequenced. The protocol developed for the isolation and sequencing of this region of the ovine protein was applied directly, and reproducibly, to bleached and unregenerated porcine and equine opsins. Comparisons of the primary structures of the fragments reveals marked variation in the sequence immediately after the lysine residue shown in the ovine protein to be the attachment point for the aldehyde group of the chromophore. Mutable positions are localized in regions previously predicted as adopting nonregular or distorted conformations and hint at structural arrangements that may provide a better understanding of the spectral and functional properties of the visual pigment.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acid Sequence Amino Acids/analysis Animals Chromatography, Gel Cyanogen Bromide Endopeptidases Horses Peptide Fragments/analysis/isolation & purification Protein Conformation Retinal Pigments Rhodopsin Serine Endopeptidases Sheep Species Specificity Swine |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > rhodopsin |
| CSHL Authors: | |
| Communities: | CSHL labs > Pappin lab |
| Depositing User: | Matt Covey |
| Date: | 1 February 1984 |
| Date Deposited: | 11 Sep 2014 14:20 |
| Last Modified: | 11 Sep 2014 14:20 |
| PMCID: | PMC1153260 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/30781 |
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