Membrane cholesterol and sphingomyelin, and ostreolysin A are obligatory for pore-formation by a MACPF/CDC-like pore-forming protein, pleurotolysin B
Membrane cholesterol and sphingomyelin, and ostreolysin A are obligatory for pore-formation by a MACPF/CDC-like pore-forming protein, pleurotolysin B
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DOI：
10.1016/j.biochi.2013.06.012
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发表时间：
2013-10-01
期刊：
影响因子：
3.9
通讯作者：
Macek, Peter
中科院分区：
文献类型：
--
作者：
Ota, Katja;Leonardi, Adrijana;Macek, Peter
平菇Pleurotus ostreatus产生溶血素(Olya)、溶胸素A(PlyA)和溶胸素B(PlyB)。目前对天然蛋白和重组蛋白的研究剖析了它们的脂结合特性以及它们在脂结合和膜通透性中的作用。利用脂质结合研究、红细胞的通透性、不同脂质成分的大单层囊泡和电子显微镜，我们发现PlyA同系物Olya优先与富含甾醇和鞘磷脂的膜结合，但不通透它们。N末端截短的Delta 48P1yB对应于天然PlyB的成熟和活性形式，它具有膜攻击复合体-穿孔素(MACPF)结构域。Delta 48PlyB在溶液中自发寡聚，并与各种脂膜弱结合，但不能穿透它们。然而，在Olya的存在下，Delta 48PlyB与胆固醇和鞘磷脂膜的结合，从而显著促进了它们的通透性。在这些膜上，Delta 48PlyB和Olya主要形成13-Meric齐聚物。它们是距膜表面9 nm厚的玫瑰花环状结构，外径19.7 nm，内径4.9 nm。当存在于相对的囊泡膜上时，这些低聚物可以二聚，从而促进囊泡的聚集。根据Delta 48PlyB的结构和功能特点，我们认为它与MACPF/胆固醇依赖的细胞溶素(CDC)蛋白有一些共同的特征。Olya是Delta 48PlyB渗透富含胆固醇和鞘磷脂的膜所必需的。(C)2013年爱思唯尔·马森公司。版权所有。
The mushroom Pleurotus ostreatus has been reported to produce the hemolytic proteins ostreolysin (OlyA), pleurotolysin A (PlyA) and pleurotolysin B (PlyB). The present study of the native and recombinant proteins dissects out their lipid-binding characteristics and their roles in lipid binding and membrane permeabilization. Using lipid-binding studies, permeabilization of erythrocytes, large unilamellar vesicles of various lipid compositions, and electron microscopy, we show that OlyA, a PlyA homolog, preferentially binds to membranes rich in sterol and sphingomyelin, but it does not permeabilize them. The N-terminally truncated Delta 48P1yB corresponds to the mature and active form of native PlyB, and it has a membrane attack complex-perforin (MACPF) domain. Delta 48PlyB spontaneously oligomerizes in solution, and binds weakly to various lipid membranes but is not able to perforate them. However, binding of Delta 48PlyB to the cholesterol and sphingomyelin membranes, and consequently, their permeabilization is dramatically promoted in the presence of OlyA. On these membranes, Delta 48PlyB and OlyA form predominantly 13-meric oligomers. These are rosette-like structures with a thickness of 9 nm from the membrane surface, with 19.7 nm and 4.9 nm outer and inner diameters, respectively. When present on opposing vesicle membranes, these oligomers can dimerize and thus promote aggregation of vesicles. Based on the structural and functional characteristics of Delta 48PlyB, we suggest that it shares some features with MACPF/cholesterol-dependent cytolysin (CDC) proteins. OlyA is obligatory for the Delta 48PlyB permeabilization of membranes rich in cholesterol and sphingomyelin. (C) 2013 Elsevier Masson SAS. All rights reserved.