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[抗菌肽盘状菌素A对细胞质膜的作用]

[Effect of discodermin A, an antimicrobial peptide, on the cytoplasm membrane].

作者信息

Sato K, Horibe K, Saito-Mitsui M, Hori M, Matsunaga S, Fusetani N, Ozaki H, Karaki H

机构信息

Department of Veterinary Pharmacology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.

出版信息

Nihon Yakurigaku Zasshi. 1997 Oct;110 Suppl 1:199P-204P. doi: 10.1254/fpj.110.supplement_199.

Abstract

The effects of discodermin A, an antimicrobial peptide extracted from sea sponge Discodermia kiiensis, on cell membranes were investigated using vascular smooth muscle cells and erythrocytes. At lower concentrations (0.1-3 microM), discodermin A increased muscle tension with an increase in intracellular free Ca2+ concentration ([Ca2+]i) indicated by the fluorescence of Ca2+ indicator, fura-PE3, in rat aortic smooth muscle. On the other hand, the higher concentration of discodermin A (10-30 microM) accelerated the leakage of loaded fura-PE3 from cells. In rabbit mesenteric artery treaded with discodermin A, addition of micromolar concentration of Ca2+ evoked contraction in the presence of ATP, suggesting that permeability of the membrane to Ca2+ and ATP is increased by discodermin A. Confocal fluorescence microscopy showed that discodermin A permeabilized the plasma membrane of A10 cells to fluorescent agents EthD-1 and the intracellular esterase coupled with another fluorescent agent calcein. Discodermin A also showed a hemolytic effect on rabbit erythrocytes, suggesting that discodermin A permitted transmembrane passage of hemoglobin. These results suggest that discodermin A form pores of different sizes on the cytoplasm membrane in concentration- and time-dependent manners. Discodermin A may be a saponin-like bioactive peptide.

摘要

使用血管平滑肌细胞和红细胞研究了从海海绵基氏盘皮海绵(Discodermia kiiensis)中提取的抗菌肽盘皮海绵素A(discodermin A)对细胞膜的影响。在较低浓度(0.1 - 3 microM)下,盘皮海绵素A增加了肌肉张力,同时大鼠主动脉平滑肌中由Ca2+指示剂fura - PE3荧光指示的细胞内游离Ca2+浓度([Ca2+]i)升高。另一方面,较高浓度的盘皮海绵素A(10 - 30 microM)加速了加载的fura - PE3从细胞中的泄漏。在用盘皮海绵素A处理的兔肠系膜动脉中,添加微摩尔浓度的Ca2+在ATP存在下引起收缩,表明盘皮海绵素A增加了膜对Ca2+和ATP的通透性。共聚焦荧光显微镜显示,盘皮海绵素A使A10细胞的质膜对荧光剂EthD - 1和与另一种荧光剂钙黄绿素偶联的细胞内酯酶通透。盘皮海绵素A对兔红细胞也有溶血作用,表明盘皮海绵素A允许血红蛋白跨膜通过。这些结果表明,盘皮海绵素A以浓度和时间依赖性方式在细胞质膜上形成不同大小的孔。盘皮海绵素A可能是一种皂苷样生物活性肽。

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