黑腹果蝇蛹中酚氧化酶原的纯化与特性分析

Purification and characterization of prophenoloxidases from pupae of Drosophila melanogaster.

作者信息

Fujimoto K, Masuda K, Asada N, Ohnishi E

机构信息

Biological Laboratory, Faculty of Science, Okayama University of Science.

出版信息

J Biochem. 1993 Mar;113(3):285-91. doi: 10.1093/oxfordjournals.jbchem.a124040.

DOI:10.1093/oxfordjournals.jbchem.a124040

PMID:8486601

Abstract

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.

摘要

从黑腹果蝇俄勒冈 - R品系的蛹中分离出两种酚氧化酶原同工型。纯化步骤包括硫酸铵分级分离、Sephacryl S - 200凝胶色谱、DEAE - 纤维素和羟基磷灰石柱色谱。这两种同工型，A1和A3，可以通过硫酸铵分级分离。A1和A3的等电点分别测定为pH 5.8和6.7。通过SDS - PAGE估计A1和A3单体的分子量分别为78 kDa和77 kDa。根据凝胶过滤色谱判断，A1和A3的天然状态被认为是同二聚体。