title: Interaction of melanosomal proteins with melanin.
abstract: Melanin is deposited in melanosomes upon a proteinaceous matrix enveloped by a melanosomal membrane. Since melanin is highly detergent insoluble, we hypothesized that the detergent solubility of proteins of the melanosomal matrix might be inversely related to the state of melanosomal melanization. Immunoblotting analyses were performed on extracts of albino and black melanocytes to test this hypothesis. The protein products of the silver (si) and the pink-eyed-dilution (p) loci as well as other matrix constituents were present at twofold higher levels in extracts of albino cells. When black cells were rendered amelanotic by growing cultures in the presence of the tyrosinase inhibitor phenylthiourea, the apparent levels of these proteins were also increased. To obviate the potential role of different levels of synthesis in contributing to these differences, we developed a cell-free melanosomal melanization assay. Upon incubation of a melanosome-rich fraction with the melanin precursor L-3,4-dihydroxyphenylalanine (Dopa) followed by immunoblot analysis, the si locus protein, the p locus protein, and other putative matrix constituents became rapidly insoluble in SDS when compared with the members of the tyrosinase-related family of melanosomal membrane proteins. Our results suggest that melanosomal proteins that interact with melanin may be identified by their relative insolubility in SDS under conditions of increasing melanization. In addition to the si locus protein and other putative melanosomal matrix proteins, the membrane-bound p locus protein may also interact closely with melanin.
authors.affiliation: Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, USA.
authors.forename: P D
authors.initials: PD
authors.lastname: Donatien
authors.forename: S J
authors.initials: SJ
authors.lastname: Orlow
mesh_terms: D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D008543:Melanins / Q000378:metabolism*; D008544:Melanocytes / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL
chemical_list: D008543:Melanins