Choi Hyemin, Hwang Jae-Sam, Lee Dong Gun
School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 702-701, Republic of Korea.
Biochim Biophys Acta. 2013 Nov;1828(11):2745-50. doi: 10.1016/j.bbamem.2013.07.021. Epub 2013 Jul 26.
The centipede Scolopendra subspinipes mutilans has been a medically important arthropod species by using it as a traditional medicine for the treatment of various diseases. In this study, we derived a novel lactoferricin B like peptide (LBLP) from the whole bodies of adult centipedes, S. s. mutilans, and investigated the antifungal effect of LBLP. LBLP exerted an antifungal and fungicidal activity without hemolysis. To investigate the antifungal mechanism of LBLP, a membrane study with propidium iodide was first conducted against Candida albicans. The result showed that LBLP caused fungal membrane permeabilization. The assays of the three dimensional flow cytometric contour plot and membrane potential further showed cell shrinkage and membrane depolarization by the membrane damage. Finally, we confirmed the membrane-active mechanism of LBLP by synthesizing model membranes, calcein and FITC-dextran loaded large unilamellar vesicles. These results showed that the antifungal effect of LBLP on membrane was due to the formation of pores with radii between 0.74nm and 1.4nm. In conclusion, this study suggests that LBLP exerts a potent antifungal activity by pore formation in the membrane, eventually leading to fungal cell death.
少棘蜈蚣作为一种用于治疗多种疾病的传统药物，一直是具有重要医学意义的节肢动物物种。在本研究中，我们从成年少棘蜈蚣的整体中获得了一种新型乳铁蛋白B样肽（LBLP），并研究了LBLP的抗真菌作用。LBLP具有抗真菌和杀菌活性且无溶血作用。为了研究LBLP的抗真菌机制，首先针对白色念珠菌进行了碘化丙啶膜研究。结果表明LBLP导致真菌细胞膜通透性增加。三维流式细胞术等高线图和膜电位分析进一步表明，膜损伤导致细胞收缩和膜去极化。最后，我们通过合成模型膜、钙黄绿素和异硫氰酸荧光素标记的葡聚糖负载的大单层囊泡，证实了LBLP的膜活性机制。这些结果表明，LBLP对膜的抗真菌作用是由于形成了半径在0.74nm至1.4nm之间的孔。总之，本研究表明LBLP通过在膜中形成孔发挥强大的抗真菌活性，最终导致真菌细胞死亡。
