IDENTIFICATION OF A MAJOR BASIC PROTEIN IN GUINEA PIG EOSINOPHIL GRANULES
IDENTIFICATION OF A MAJOR BASIC PROTEIN IN GUINEA PIG EOSINOPHIL GRANULES
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豚鼠嗜酸性粒细胞颗粒中主要碱性蛋白的鉴定
DOI：
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发表时间：
1973
影响因子：
15.3
通讯作者：
J. Maldonado
中科院分区：
文献类型：
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作者：
G. Gleich;D. Loegering;J. Maldonado
嗜酸性粒细胞功能的阐明可通过颗粒成分的分析来完成。我们已经从豚鼠腹腔中纯化了嗜酸性粒细胞（93%或更高），并研究了各种方法来破坏细胞并释放完整的颗粒。在0.34 M蔗糖裂解得到最好的产量的颗粒，这些具有特征性的形态嗜酸性粒细胞颗粒时，通过电子显微镜检查。通过各种处理溶解颗粒，并通过在pH 3的6 M尿素中的聚丙烯酰胺电泳分析溶液。溶解的嗜酸性粒细胞和中性粒细胞颗粒的电泳图谱的比较显示了差异：来自嗜酸性粒细胞颗粒的蛋白质的主要部分（53±3%; x ±1 SE）作为单一组分迁移。这种主带蛋白的分子量在6，000和12，000道尔顿之间，pI为10或更大。Sephadex G-50上嗜酸性粒细胞颗粒成分的分析显示两个主峰;峰1具有过氧化物酶活性，峰2含有主带蛋白。这些研究表明，嗜酸性粒细胞颗粒含有低分子量的阳离子蛋白，其缺乏过氧化物酶活性，占颗粒蛋白的50%以上。
Elucidation of the functions of the eosinophil might be accomplished by analysis of the granule constituents. We have purified eosinophils (93% or greater) from the peritoneal cavity of the guinea pig and have investigated a variety of methods to disrupt cells and liberate intact granules. Lysis in 0.34 M sucrose gave the best yield of granules and these had the characteristic morphology of eosinophil granules when examined by electron microscopy. Granules were solubilized by a variety of treatments and the solutions analyzed by polyacrylamide electrophoresis at pH 3 in 6 M urea. Comparison of the electrophoretic patterns of solubilized eosinophil and neutrophil granules revealed a difference: a major portion (53±3%; x ±1 SE) of the protein from the eosinophil granule migrated as a single component. This major band protein has a molecular weight between 6,000 and 12,000 daltons and a pI of 10 or greater. Analysis of eosinophil granule constituents on Sephadex G-50 revealed two main peaks; peak 1 possessed peroxidase activity and peak 2 contained the major band protein. These studies indicate that eosinophil granules contain a cationic protein of low molecular weight which lacks peroxidase activity and which accounts for greater than 50% of granule protein.