Modrzakowski M C, Paranavitana C M
Infect Immun. 1981 May;32(2):668-74. doi: 10.1128/iai.32.2.668-674.1981.
Granule contents from human polymorphonuclear leukocytes were prepared by extraction with 0.2 M acetate, pH 4. A buffer extract fraction (peak D) obtained by Sephadex G-100 column chromatography demonstrated distinct antimicrobial activity toward Acinetobacte sp. independent of added H2O2 or Cl-. The protein of this fraction had an apparent molecular weight of 9,000 and demonstrated time and dose dependence that was more active against stationary-growth cells than mid-log-phase cells. The bactericidal activity of the fraction was most active at 37 degrees C, with only slight activity demonstrated at 22 degrees C and no activity at 4 degrees C. Boiling the granule fraction for 30 min did not affect the antimicrobial activity. However, pronase or trypsin pretreatment of the peak D fraction reduced its antimicrobial activity. When the membrane lipid composition of Acinetobacter sp. was altered by growth on specific n-alkane carbon sources, the susceptibility to the granule fraction was also altered. Resistance to the activity of the granule fraction increased as the carbon chain length of the growth substrate increased. Liposomes formed from Acinetobacter sp. lipid extracts and containing glucose were made leaky with the addition of the granule fraction (boiled and not boiled), suggesting a membrane-disruptive activity of the granule protein against Acinetobacter sp. membranes.
通过用pH 4的0.2M醋酸盐提取制备人多形核白细胞的颗粒内容物。通过Sephadex G - 100柱色谱获得的缓冲提取物级分（峰D）对不动杆菌属表现出明显的抗菌活性，且与添加的H2O2或Cl-无关。该级分的蛋白质表观分子量为9000，表现出时间和剂量依赖性，对静止期生长细胞的活性比对对数中期细胞的活性更高。该级分的杀菌活性在37℃时最活跃，在22℃时仅表现出轻微活性，在4℃时无活性。将颗粒级分煮沸30分钟不影响抗菌活性。然而，用链霉蛋白酶或胰蛋白酶预处理峰D级分会降低其抗菌活性。当不动杆菌属的膜脂质组成通过在特定正构烷烃碳源上生长而改变时，对颗粒级分的敏感性也会改变。随着生长底物碳链长度的增加，对颗粒级分活性的抗性增加。由不动杆菌属脂质提取物形成并含有葡萄糖的脂质体在添加颗粒级分（煮沸和未煮沸）后变得渗漏，表明颗粒蛋白对不动杆菌属膜具有膜破坏活性。
