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A hypothesis as to mechanism of perturbation in biomembrane due to endotoxin. Participation of lysosomal cationic proteins with an endotoxin binding capacity

关于内毒素引起的生物膜扰动机制的假设。

基本信息

批准号：
60480167
负责人：
YOSHIDA Masao
金额：
$ 4.03万
依托单位：
Ieate Medical University, School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

项目摘要

1) Fractionation of cationic proteins (CAP's); CAP's were fractionated from rebbit peritoneal granulocytes through the steps of 0.1 M citric acid extraction, ethanol precipitation and gel filtration by FPLC. Since CAP's were capable of binding with heparin, a purification using heparin Sepharose is being investigated.2) CAP's agglutinated the erythrocytes sensitized with LPS, especially Re-LPS (CAP-HA); CAP-HA was inhibited by preincubation of CAP with LPS.3) CAP bound with LPS, especially Re-LPS in water solution, and made increased turbidity; the increase of turbidity depended upon CAP-levels, although the binding took place at an optimal ratio of both CAP and LPS levels; Low ionic strength condition and pH 6-7 were optimal to CAP-HA; the binding was reversible. This seems to be involved in both ionic and hydrophobic bounds of CAP and LPS. CAP-HA is thought to result from the binding mentioned above.4) CAP exhibited antibacterial activity against S and R form gram-negative bacteria as well as gram-positive bacteria, expecially against R from gram-negatives. The activity seems to be bacteriolytic.5) CAP bound with heparin made increased trubidity, consequently the binding neutralized mutually the properties of CAP and heparin.6) One CAP fraction of less than 10,000 M.W. had high anticoagulant activity compared to other properties.7) It seems that CAP can promote the activity of LPS, which bound with cells such as erythrocytes, and membrane perturbation.

1)阳离子蛋白的分离；经0.1 M柠檬酸提取、乙醇沉淀、高效液相色谱凝胶过滤，分离兔腹膜粒细胞中CAP。由于CAP能够与肝素结合，目前正在研究使用肝素Sepharose进行纯化。2) CAP对LPS致敏的红细胞，尤其是Re-LPS （CAP- ha）有凝集作用；3) CAP与LPS结合，尤其是在水溶液中与Re-LPS结合，使其浊度增加；浊度的增加依赖于CAP水平，尽管结合在CAP和LPS水平的最佳比例下发生；低离子强度条件和pH 6 ~ 7对CAP-HA最有利；装订是可逆的。这似乎涉及到CAP和LPS的离子和疏水界。CAP-HA被认为是上述结合的结果。4) CAP对革兰氏阴性菌S、R及革兰氏阳性菌均表现出抑菌活性，对革兰氏阴性菌R的抑菌活性尤其明显。这种活性似乎具有溶菌作用。5) CAP与肝素结合使其浊度增加，从而使CAP与肝素的特性相互中和。6)与其他性能相比，小于10,000 m.w的CAP组分具有较高的抗凝活性。7) CAP似乎可以促进与红细胞等细胞结合的LPS活性和膜摄动。