Subcellular Localization and Heterogeneity of Neutral Proteases in Neutrophilic Polymorphonuclear Leukocytes
Subcellular Localization and Heterogeneity of Neutral Proteases in Neutrophilic Polymorphonuclear Leukocytes
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中性粒细胞多形核白细胞中性蛋白酶的亚细胞定位和异质性
DOI：
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发表时间：
1975
影响因子：
15.3
通讯作者：
M. Baggiolini
中科院分区：
文献类型：
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作者：
B. Dewald;R. Rindler‐Ludwig;U. Bretz;M. Baggiolini
用等密度离心法对人和兔多形核白细胞中弹性蛋白酶和水解组蛋白和酪蛋白的中性蛋白酶的亚细胞定位进行了研究。以N-乙酰-D、L-丙氨酸α-萘酯或萘酚AS-D醋酸酯为底物进行染色，验证了凝胶的蛋白水解性。在这两个物种中，所有被检测的中性蛋白水解酶都只定位在天青颗粒中。人的比活力约为兔制剂的10-30倍。在人嗜天青颗粒提取物中，经电泳法分离后，可鉴定出多达10种具有酯解活性的蛋白质。这些酯酶的三个主要组分和两到三个次要组分具有弹性淀粉酶活性。用兔嗜天青颗粒提取液制备的类似的酶谱显示只有一条主要的弹性酶带。进一步的电泳分析表明，人和兔中性粒细胞中最强的阳离子蛋白也局限于天青颗粒。
The subcellular localization of elastase and of neutral proteases hydrolyzing histone and casein was determined in human and rabbit polymorphonuclear leukocytes using fractionation by isopycnic centrifugation. Granule-rich fractions obtained by this technique were extracted and analyzed by acrylamide gel electrophoresis, and proteolytic activity on the gels was demonstrated by staining with either N-acetyl-D,L-alanine alpha-naphthyl ester or naphthol AS-D acetate as substrate. In both species, all neutral proteases assayed were found to be localized exclusively in the azurophil granules. Specific activities were about 10-30 times higher in human than in rabbit preparations. In extracts of human azurophil granules up to 10 proteins exhibiting esterolytic activity could be demonstrated after electrophoretic separation. Three major and two or three minor components of these esterases were shown to possess elastase activity. Similar zymograms prepared with extracts from rabbit azurophil granules revealed only one major elastase band. The electrophoretic analysis further showed that the most strongly cationic proteins of both human and rabbit PMNs were also confined to the azurophil granules.