title: Bioactive motifs of agouti signal protein.
abstract: The switch between the synthesis of eu- and pheomelanins is modulated by the interaction of two paracrine signaling molecules, alpha-melanocyte stimulating hormone (MSH) and agouti signal protein (ASP), which interact with melanocytes via the MSH receptor (MC1R). Comparison of the primary sequence of ASP with the known MSH pharmacophore provides no suggestion about the putative bioactive domain(s) of ASP. To identify such bioactive motif(s), we synthesized 15-mer peptides that spanned the primary sequence of ASP and determined their effects on the melanogenic activities of murine melanocytes. Northern and Western blotting were used, together with chemical analysis of melanins and enzymatic assays, to identify three distinct bioactive regions of ASP that down-regulate eumelanogenesis. The decrease in eumelanin production was mediated by down-regulation of mRNA levels for tyrosinase and other melanogenic enzymes, as occurs in vivo, and these effects were comparable to those elicited by intact recombinant ASP. Shorter peptides in those motifs were synthesized and their effects on melanogenesis were further investigated. The amino acid arginine, which is present in the MSH peptide pharmacophore (HFRW), is also in the most active domain of ASP (KVARP). Our data suggest that lysines and an arginine (in motifs such as KxxxxKxxR or KxxRxxxxK) are important for the bioactivity of ASP. Identification of the specific ASP epitope that interacts with the MC1R has potential pharmacological applications in treating dysfunctions of skin pigmentation.
authors.affiliation: Laboratory of Cell Biology, National Cancer Institute, Bethesda, Maryland, 20892, USA.
authors.forename: V M
authors.initials: VM
authors.lastname: Virador
authors.forename: C
authors.initials: C
authors.lastname: Santis
authors.forename: M
authors.initials: M
authors.lastname: Furumura
authors.forename: H
authors.initials: H
authors.lastname: Kalbacher
authors.forename: V J
authors.initials: VJ
authors.lastname: Hearing
mesh_terms: D054366:Agouti Signaling Protein; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D008543:Melanins / Q000096:biosynthesis; D008544:Melanocytes / Q000737:chemistry / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000235:genetics; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D011979:Receptors, Pituitary Hormone / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D000521:alpha-MSH / Q000494:pharmacology
chemical_list: D054366:Agouti Signaling Protein; D036341:Intercellular Signaling Peptides and Proteins; D008543:Melanins; D010446:Peptide Fragments; D011506:Proteins; D012333:RNA, Messenger; D011979:Receptors, Pituitary Hormone; D000521:alpha-MSH; C029446:MSH receptor; D000242:Cyclic AMP; D014442:Monophenol Monooxygenase; D019204:GTP-Binding Proteins