title: Complex formation between mushroom tyrosinase and Manduca dopachrome isomerase.
abstract: Melanin biosynthesis in animals is initiated by the ubiquitously present tyrosinase and is aided by dopachrome isomerase. We have characterized a novel dopachrome isomerase (decarboxylating) from the hemolymph of Manduca sexta that generates a new quinone methide intermediate during melanogenesis (Sugumaran, M. and Semensi, V. (1991) J. Biol. Chem. 266, 6073-6078). This enzyme has the ability to form a complex with mushroom tyrosinase as judged by a number of physicochemical studies. The isomerase exhibited a marked inhibitory effect on tyrosinase and tyrosinase reciprocated by inhibiting the isomerase. While the isomerase showed no activity toward preformed dopaminechrome, it readily influenced the stability of dopaminechrome generated in situ by tyrosinase. Moreover, mushroom tyrosinase, which lacked specific binding to Concanavalin A Sepharose column, after complexing with the isomerase exhibited binding to this column. The complex formation also affected the pI value as well as mobility on a size exclusion column of these enzymes. Enzymes executing sequential metabolic transformation are known to form complexes called metabolons. Based on these above studies, it is concluded that both the enzymes involved in insect melanogenic pathway--phenoloxidase and dopachrome isomerase--are able to form a metabolon complex.
authors.affiliation: Department of Biology, University of Massachusetts at Boston 02125, USA.
authors.forename: M
authors.initials: M
authors.lastname: Sugumaran
authors.forename: K
authors.initials: K
authors.lastname: Nellaiappan
authors.forename: T
authors.initials: T
authors.lastname: Scott
authors.forename: C
authors.initials: C
authors.lastname: Amaratunga
mesh_terms: D000818:Animals; D001487:Basidiomycota / Q000201:enzymology*; D002845:Chromatography; D003208:Concanavalin A; D004795:Enzyme Stability; D019746:Intramolecular Oxidoreductases; D007525:Isoelectric Focusing; D007535:Isomerases / Q000032:analysis / Q000378:metabolism*; D007700:Kinetics; D018402:Manduca / Q000201:enzymology*; D008543:Melanins / Q000096:biosynthesis*; D008970:Molecular Weight; D014442:Monophenol Monooxygenase / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding; D012685:Sepharose
chemical_list: D008543:Melanins; D003208:Concanavalin A; D012685:Sepharose; D014442:Monophenol Monooxygenase; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase