Purification and characterization of prophenoloxidases from pupae of Drosophila melanogaster.
Two isoforms of prophenoloxidase were isolated from pupae of Oregon-R strain of Drosophila melanogaster. The purification procedure included ammonium sulfate fractionation, Sephacryl S-200 gel chromatography, DEAE-cellulose, and hydroxylapatite column chromatography. The two isoforms, A1 and A3, could be separated by ammonium sulfate fractionation. The isoelectric points of A1 and A3 were determined to be pH 5.8 and 6.7, respectively. The molecular weights of the monomers of A1 and A3 were estimated by SDS-PAGE to be 78 and 77 kDa, respectively. The native states of A1 and A3 are considered to be homodimeric, as judged by gel-filtration chromatography.
Fujimoto
K
K
Biological Laboratory, Faculty of Science, Okayama University of Science.
Masuda
K
K
Asada
N
N
Ohnishi
E
E
D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D004156:Catechol Oxidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D004331:Drosophila melanogaster / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D011679:Pupa / Q000201:enzymology; D013379:Substrate Specificity; D013696:Temperature
D004792:Enzyme Precursors; D007527:Isoenzymes; C038451:pro-phenoloxidase; D004156:Catechol Oxidase; D000645:Ammonium Sulfate