人白细胞髓过氧化物酶的纯化及鉴定

Purification and Identification of Human Leukocyte Myeloperoxidase

以十六烷基三甲基溴化铵溶液提取正常人白细胞髓过氧化物酶经硫酸铵分级沉淀,过SephadexG-150柱,获得其纯品,RZ值为0.68,酶活性为29.77u/mg,SDS-PAGE谱显示3条区带,分子量分别为59kd、38kd、13.5kd。

Myeloperoxidase (MPO)plays an impor-tant role in the oxygen--dependent micro-bicidal mechanism of polymorphonuclearneutrophils. The purpose of our study wasto investigate the therapeutic potency ofhuman MPO preparations. This paper isto present our work on MPO purificationand its identification. White blood cells, isolated freshly fromnormal donors, were lysed with cetyl-trimethylammonium bromide to liberatemyeloperoxidase. The purified MPO wasobtained by 65％ (NH_4)_2SO_4 precipita-tion followed by separation over theSephadex G150 column. These pure MPOpreparations were green in color and hadan A_(430)/A_(230) nm of 0.68. The enzymaticactivity was of 29.77u/mg. The pure nor-mal MPO was composed of a 59 000MWpeptide, a 13 500MW peptide. and a 38 000MW peptide when subjected to SDS--PAGEunder reducing conditions. The study of the therapeutic effect ofthe pure MPO on mice with C. albicans in-fection is under way.