title: Crithidia guilhermei: purification and partial characterization of a 62-kDa extracellular metalloproteinase.
abstract: An extracellular metalloproteinase from Crithidia guilhermei, a monoxenic trypanosomatid of insects, was purified 11-fold by ammonium sulfate precipitation, gel filtration on a Shinpack Diol-150 column, and anion-exchange chromatography in a MONO Q column, both using the HPLC system. The proteinase appeared as a single band with an apparent molecular mass of 62 kDa in SDS-PAGE, under reducing conditions, and was optimally active at 37 degrees C and pH 6.0. The enzyme showed 62% residual activity at 50 degrees C for 30 min. The proteinase was completely inhibited by 1, 10-phenanthroline, indicating that the enzyme belongs to the metalloproteinase class. This is the first report of the purification of an extracellular metalloproteinase from the Crithidia species. The possible role of this enzyme in the digestive tract of the insect host is discussed.
authors.affiliation: Department of General Microbiology, Institute of Microbiology Prof Paulo de Góoes, Rio de Janeiro, RJ, 21941-590, Brazil.
authors.forename: A C
authors.initials: AC
authors.lastname: Nogueira de Melo
authors.forename: S
authors.initials: S
authors.lastname: Giovanni-De-Simone
authors.forename: M H
authors.initials: MH
authors.lastname: Branquinha
authors.forename: A B
authors.initials: AB
authors.lastname: Vermelho
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003421:Crithidia / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007313:Insecta; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification*; D008970:Molecular Weight; D010618:Phenanthrolines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D013696:Temperature; D015032:Zinc / Q000494:pharmacology
chemical_list: D010618:Phenanthrolines; D011480:Protease Inhibitors; D008666:Metalloendopeptidases; D015032:Zinc; C025205:1,10-phenanthroline