title: Characterization of a new phenoloxidase inhibitor from the cuticle of Manduca sexta.
abstract: Melanin, the phenolic biopolymer that serves as a skin- and hair pigment-protecting agent against harmful solar radiation and a free radical trap, is biosynthesized in animals mainly by the action of tyrosinase also known as phenoloxidase. Regulation of tyrosinase and hence melanogenesis is vital for all animals. In this report, we present the isolation and characterization of a new, heat-labile glycoprotein inhibitor of phenoloxidase from the larvae of Manduca sexta. The inhibitor was isolated from the live larval cuticle by buffer extraction and purified to homogeneity employing ammonium sulfate precipitation, dialysis, and concanavalin A-Sepharose chromatography. It migrated with a molecular weight of 380,000 on SDS-PAGE gels and inhibited the activity of insect and plant as well as fungal phenoloxidases. Inhibitor formed a tight complex with phenoloxidases, which resisted dissociation even by 1% Triton X-100 or SDS. Selective inhibition of phenoloxidase, while acting on certain but not all different substrates, was observed. The physiological importance of this newly discovered high-molecular-weight phenoloxidase inhibitor is discussed.
authors.affiliation: Department of Biology, University of Massachusetts Boston, 100 Morrissey Boulevard, Boston, Massachusetts 02125, USA. manickam.sugumaran@umb.edu
authors.forename: M
authors.initials: M
authors.lastname: Sugumaran
authors.forename: K
authors.initials: K
authors.lastname: Nellaiappan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D007814:Larva / Q000737:chemistry; D018402:Manduca / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors*; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D017421:Sequence Analysis
chemical_list: D004791:Enzyme Inhibitors; D010455:Peptides; D014442:Monophenol Monooxygenase