GUINEA-PIG HETEROPHIL AND EOSINOPHIL PEROXIDASE
GUINEA-PIG HETEROPHIL AND EOSINOPHIL PEROXIDASE
复制标题
DOI：
10.1016/0003-9861(72)90164-6
复制
发表时间：
1972-01-01
影响因子：
3.9
通讯作者：
HIMMELHO.R
中科院分区：
文献类型：
--
作者：
DESSER, RK;EVANS, WH;HIMMELHO.R
为了比较某些物理性质，从骨髓白细胞中纯化了豚鼠嗜异粒细胞和嗜酸性粒细胞过氧化物酶。纯化的嗜异性过氧化物酶（通过免疫学试验和凝胶电泳在超电泳中均质）在437 nm处具有Soret最大值，A 437 A 280比率为0.79，mol wt为137，000。该分子由两个大小相似的亚基组成，如通过其在电子显微镜下的外观和在SDS聚丙烯酰胺凝胶中的电泳所评估。嗜酸性粒细胞过氧化物酶需要阳离子洗涤剂十六烷基三甲基溴化铵，以防止结合到其纯化中使用的材料。纯化的嗜酸性粒细胞酶（根据免疫学标准是同质的）通过凝胶过滤分解为两种过氧化物酶组分，分子量分别为75，000和150，000。各组分具有相同的比活性、吸收光谱（Soret最大值位于425 nm处）和A 415 A 280比值（0.83）。用SDS和巯基乙醇处理每一种产生摩尔重量约65，000的单一组分。对骨髓粗提物的检查仅分离出一种嗜酸性粒细胞过氧化物酶组分，分子量约为80，000，表明在纯化过程中发生了聚集。因此，尽管白细胞过氧化物酶位于密切相关细胞的类似细胞器中，但它们具有实质上不同的性质。
Guinea pig heterophil and eosinophil peroxidases were purified from bone marrow leukocytes in order to compare certain physical properties. Purified heterophil peroxidase (homogeneous in the ultracentrifuge, by immunologic test, and by gel electrophoresis) had a Soret maximum at 437 nm, an A 437 A 280 ratio of 0.79, and a mol wt of 137,000. The molecule consisted of two subunits of similar size, as assessed by its appearance in the electron microscope and by electrophoresis in SDS polyacrylamide gels. Eosinophil peroxidase required the cationic detergent cetyltrimethylammonium bromide to prevent binding to the materials used in its purification. The purified esoinophil enzyme, homogeneous by immunologic criteria, was resolved by gel filtration into two peroxidase components having mol wts of 75,000 and 150,000, respectively. Each component had the same specific activity, absorption spectrum (Soret maximum at 425 nm), and A 415 A 280 ratio (0.83). Treatment of each with SDS and mercaptoethanol produced a single component with a mol wt of about 65,000. Examination of crude bone marrow extracts resolved only one eosinophil peroxidase component with a mol wt of about 80,000, indicating that aggregation occurred during purification. The leukocyte peroxidases, therefore, have substantially different properties despite their location in analogous organelles of closely related cells.