title: Mass spectrometric analysis of porcine rhodopsin.
abstract: Rhodopsin is the dim light photosensitive pigment of animals. In this work, we undertook to study the structure of rhodopsin from swine and compare it with bovine and rat rhodopsin. Porcine rhodopsin was analyzed using methodology developed previously for mass spectrometric analysis of integral membrane proteins. Combining efficient protein cleavage and high performance liquid chromatography separation with the sensitivity of mass spectrometry (MS), this technique allows the observation of the full protein map and the posttranslational modifications of the protein in a single experiment. The rhodopsin protein from a single porcine eye was sequenced completely, with the exception of two single-amino acid fragments and one two-amino acid fragment, and the gene sequence reported previously was confirmed. The posttranslational modifications, similar to the ones reported previously for bovine and rat rhodopsin, were also identified. Although porcine rhodopsin has a high degree of homology to bovine and rat rhodopsins and most of their posttranslational modifications are identical, the glycosylation and phosphorylation patterns observed were different. These results show that rhodopsin from a single porcine eye can be characterized completely by MS. This technology opens the possibility of rhodopsin structural and functional studies aided by powerful mass spectrometric analysis, using the fellow eye as an internal control.
authors.affiliation: Department of Ophthalmology, Medical University of South Carolina, Charleston 29425, USA.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Ablonczy
authors.forename: Patrice
authors.initials: P
authors.lastname: Goletz
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Knapp
authors.forename: Rosalie K
authors.initials: RK
authors.lastname: Crouch
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D051381:Rats; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism; D013552:Swine
chemical_list: D012243:Rhodopsin