Cooray R, Petersson C G, Holmberg O
Department of Veterinary Medical Microbiology, Swedish University of Agricultural Sciences, Uppsala.
Vet Immunol Immunopathol. 1993 Oct;38(3-4):261-72. doi: 10.1016/0165-2427(93)90086-j.
Bovine myeloperoxidase (MPO) was isolated and purified from neutrophil granules using protein extraction at pH 4 and gel filtration combined with fast protein liquid chromatography. The extracted protein was identified as MPO based on its absorption spectrum, amino acid composition, peroxidase activity and polypeptide structure. Bovine neutrophils contained three different forms of MPO (I, II and III). When subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis each of the three purified forms showed two distinct bands corresponding to heavy and light polypeptide chains of approximately 57,000 and 15,000 molecular radius. Amino acid analysis of the three forms showed that there was an overall similarity between them. Slight differences were found between MPO Form III and the other two forms. The three forms of bovine MPO were shown to differ in their specific enzyme activities in a luminol-dependent chemiluminescence assay. MPO Form III showed the highest enzyme activity. The amount recovered during purification of the respective MPO forms varied, with the recovery being highest for MPO I. Our findings suggest that there are intrinsic differences between the three forms of bovine MPO. In terms of their amino acid composition and molecular weight, the bovine MPO closely resembled human and canine MPO.
使用pH 4的蛋白质提取方法，结合凝胶过滤和快速蛋白质液相色谱法，从嗜中性粒细胞颗粒中分离并纯化牛髓过氧化物酶（MPO）。根据其吸收光谱、氨基酸组成、过氧化物酶活性和多肽结构，将提取的蛋白质鉴定为MPO。牛嗜中性粒细胞含有三种不同形式的MPO（I、II和III）。当进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳时，三种纯化形式中的每一种都显示出两条明显的条带，分别对应于分子量约为57,000和15,000的重链和轻链多肽。对这三种形式的氨基酸分析表明，它们之间总体相似。在MPO III型与其他两种形式之间发现了细微差异。在鲁米诺依赖性化学发光测定中，三种形式的牛MPO在其特定酶活性方面表现出差异。MPO III型显示出最高的酶活性。纯化过程中各MPO形式的回收量各不相同，其中MPO I的回收率最高。我们的研究结果表明，三种形式的牛MPO之间存在内在差异。就氨基酸组成和分子量而言，牛MPO与人和犬MPO非常相似。
