title: Proteomic identification of unique photoreceptor disc components reveals the presence of PRCD, a protein linked to retinal degeneration.
abstract: Visual signal transduction takes place on the surface of flat membrane vesicles called photoreceptor discs, which reside inside the light-sensitive outer segment organelle of vertebrate photoreceptor cells. Although biochemical studies have indicated that discs are built with a handful of highly specialized proteins, proteomic studies have yielded databases consisting of hundreds of entries. We addressed this controversy by employing protein correlation profiling, which allows identification of unique components of organelles that can be fractionated but not purified to absolute homogeneity. We subjected discs to sequential steps of fractionation and identified the relative amounts of proteins in each fraction by label-free quantitative mass spectrometry. This analysis demonstrated that the photoreceptor disc proteome contains only eleven components, which satisfy the hallmark criterion for being unique disc-resident components: the retention of a constant molar ratio among themselves across fractionation steps. Remarkably, one of them is PRCD, a protein whose mutations have been shown to cause blindness, yet cellular localization remained completely unknown. Identification of PRCD as a novel disc-specific protein facilitates understanding its functional role and the pathobiological significance of its mutations. Our study provides a striking example how protein correlation profiling allows a distinction between constitutive components of cellular organelles and their inevitable contaminants.
authors.affiliation: Albert Eye Research Institute, 2Institute for Genome Sciences & Policy, Duke University School of Medicine, Durham, North Carolina 27710, United States.
authors.forename: Nikolai P
authors.initials: NP
authors.lastname: Skiba
authors.forename: William J
authors.initials: WJ
authors.lastname: Spencer
authors.forename: Raquel Y
authors.initials: RY
authors.lastname: Salinas
authors.forename: Eric C
authors.initials: EC
authors.lastname: Lieu
authors.forename: J Will
authors.initials: JW
authors.lastname: Thompson
authors.forename: Vadim Y
authors.initials: VY
authors.lastname: Arshavsky
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002458:Cell Fractionation; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009154:Mutation; D040901:Proteomics; D012162:Retinal Degeneration / Q000235:genetics* / Q000473:pathology; D055214:Retinal Photoreceptor Cell Outer Segment / Q000737:chemistry* / Q000378:metabolism
chemical_list: D005136:Eye Proteins; C583558:PRCD protein, human