Sato K, Horibe K, Saito-Mitsui M, Hori M, Matsunaga S, Fusetani N, Ozaki H, Karaki H
Department of Veterinary Pharmacology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.
Nihon Yakurigaku Zasshi. 1997 Oct;110 Suppl 1:199P-204P. doi: 10.1254/fpj.110.supplement_199.
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.
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。