Potency of bactericidal proteins purified from the large granules of bovine neutrophils
Potency of bactericidal proteins purified from the large granules of bovine neutrophils
复制标题
从牛中性粒细胞大颗粒中纯化的杀菌蛋白的效力
作者：
R. Gennaro;L. Dolzani;D. Romeo
牛中性粒细胞的大颗粒的新的人口，这是独立的O2衍生物的杀菌活性的细胞库，提取与酸性介质。通过离子交换色谱法（用羧甲基纤维素）和凝胶过滤（用Sephadex G-50）从提取物中分离出几个馏分。这些馏分中的一些只含有非常有限的主要成分，通过聚丙烯酰胺凝胶电泳检测。在含有生理浓度Na+、K+、Mg 2+和Ca 2+的培养基中，纯化的杀菌蛋白在0.1至10微克/0.3 ml含有1 × 10（6）至2 × 10（6）CFU金黄色葡萄球菌或大肠杆菌的测定混合物中显示出其活性。两个组分，含有多肽的表观分子量范围从28，000到小于12，000，引起相当选择性和快速（5至20分钟）杀死S。金黄色。它们的作用伴随着一些低分子量（小于12，000）组分与革兰氏阳性菌的显著结合。其他SephadexG-50组分，含有相对高分子量的第一个出现的蛋白质，对E. coli比S.金黄色。对于革兰氏阴性菌，杀菌活性的开始有10分钟的延迟，此后发展非常快。在体外效力的大颗粒杀菌蛋白的基础上，我们建议，即使部分放电的颗粒内容物进入吞噬体可以提供一个非常有效的非氧化杀菌系统作用于革兰氏阳性和革兰氏阴性微生物的牛嗜中性粒细胞的吞噬空泡。
The novel population of large granules of bovine neutrophils, which is the cell store of bactericidal activity independent of O2 derivatives, was extracted with an acid medium. Several fractions were resolved from the extract by ion-exchange chromatography (with carboxymethyl-cellulose) and gel filtration (with Sephadex G-50). Some of these fractions contained only a very limited number of major components, as detected by polyacrylamide gel electrophoresis. The purified bactericidal proteins exhibited their activity at 0.1 to 10 micrograms/0.3 ml of assay mixture containing 1 X 10(6) to 2 X 10(6) CFU of Staphylococcus aureus or Escherichia coli in media with physiological concentrations of Na+, K+, Mg2+, and Ca2+. Two fractions, containing polypeptides with apparent molecular weights ranging from 28,000 to less than 12,000, caused rather selective and rapid (5 to 20 min) killing of S. aureus. Their action was accompanied by significant binding to the gram-positive bacteria of some low (less than 12,000)-molecular-weight components. Other Sephadex G-50 fractions, containing the first emerging proteins with relatively high molecular weights, were more active on E. coli than on S. aureus. With the gram-negative bacteria there was a 10-min delay in the onset of bactericidal activity, which thereafter developed very fast. On the basis of the in vitro potency of the large-granule bactericidal proteins, we suggest that even partial discharge of granule content into the phagosomes may supply the phagocytic vacuoles of bovine neutrophils with a very efficient nonoxidative bactericidal system acting on both gram-positive and gram-negative microorganisms.