Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia; BioSistemika Ltd, Ljubljana, Slovenia.
Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.
Biochim Biophys Acta Biomembr. 2020 Sep 1;1862(9):183307. doi: 10.1016/j.bbamem.2020.183307. Epub 2020 Apr 13.
Oyster mushrooms (Pleurotus spp.) have recently been shown to produce insecticidal bi-component protein complexes based on the aegerolysin proteins. A role for these proteins is thus indicated for defence and protection of the mushroom, and we propose their use as new environmentally friendly bioinsecticides. These aegerolysin-based protein complexes permeabilise artificial lipid vesicles through aegerolysin binding to an insect-specific sphingolipid, ceramide phosphoethanolamine (CPE), and they are cytotoxic for the Spodoptera frugiferda (Sf9) insect cell line. Tandem mass spectrometry analysis of the Sf9 lipidome uncovered lipids not previously reported in the literature, including in particular C14 sphingosine-based CPE molecular species, which comprised ~4 mol% of the whole lipidome. Further analysis of the lipid binding specificity of an aegerolysin from P. ostreatus, ostreolysin A6 (OlyA6), to lipid vesicles composed of commercial lipids, to lipid vesicles composed of the total lipid extract from Sf9 cells, and to HPLC-separated Sf9 cell lipid fractions containing ceramides, confirmed CPE as the main OlyA6 receptor, but also highlighted the importance of membrane cholesterol for formation of strong and stable interactions of OlyA6 with artificial and natural lipid membranes. Binding assays performed with glycan arrays and surface plasmon resonance, which included invertebrate-specific glycans, excluded these saccharides as potential additional OlyA6 receptors.
蚝蘑（Pleurotus spp.）最近被发现能够产生基于蜂毒素蛋白的杀虫双组分蛋白复合物。这些蛋白在蘑菇的防御和保护中具有重要作用，因此我们提议将其用作新型环保的生物杀虫剂。这些基于蜂毒素的蛋白复合物通过蜂毒素与昆虫特异性神经酰胺磷酸乙醇胺（CPE）结合来渗透人工脂质囊泡，并且对 Spodoptera frugiferda（Sf9）昆虫细胞系具有细胞毒性。对 Sf9 脂质组的串联质谱分析揭示了以前文献中未报道的脂质，特别是基于 C14 神经酰胺的 CPE 分子种类，其占整个脂质组的约 4 mol%。进一步分析来自 P. ostreatus 的蜂毒素 ostreolysin A6（OlyA6）与由商业脂质组成的脂质囊泡、与 Sf9 细胞总脂质提取物组成的脂质囊泡以及与包含神经酰胺的 HPLC 分离 Sf9 细胞脂质级分的脂质结合特异性，证实 CPE 是 OlyA6 的主要受体，但也强调了膜胆固醇对 OlyA6 与人工和天然脂质膜形成强而稳定相互作用的重要性。用糖缀合物阵列和表面等离子体共振进行的结合测定，其中包括无脊椎动物特异性聚糖，排除了这些糖作为潜在的额外 OlyA6 受体。
