Zaruba Marianne, Roschitz Lena, Sami Haider, Ogris Manfred, Gerner Wilhelm, Metzner Christoph
Institute of Virology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria.
Laboratory of Macromolecular Cancer Therapeutics MMCT, Department of Pharmaceutical Sciences, University of Vienna, 1090 Vienna, Austria.
Membranes (Basel). 2021 Jun 4;11(6):428. doi: 10.3390/membranes11060428.
Extracellular vesicles produced by different types of cells have recently attracted great attention, not only for their role in physiology and pathology, but also because of the emerging applications in gene therapy, vaccine production and diagnostics. Less well known than their eukaryotic counterpart, also bacteria produce extracellular vesicles, in the case of the Gram-negative the main species is termed outer membrane vesicles (OMVs). In this study, we show for the first time the functional surface modification of OMVs with glycosylphosphatidylinositol (GPI)-anchored protein, exploiting a process variably described as molecular painting or protein engineering in eukaryotic membranes, whereby the lipid part of the GPI anchor inserts in cell membranes. By transferring the process to bacterial vesicles, we can generate a hybrid of perfectly eukaryotic proteins (in terms of folding and post-translational modifications) on a prokaryotic platform. We could demonstrate that two different GPI proteins can be displayed on the same OMV. In addition to fluorescent marker proteins, cytokines, growth factors and antigens canb be potentially transferred, generating a versatile modular platform for a novel vaccine strategy.
不同类型细胞产生的细胞外囊泡最近备受关注，这不仅是因为它们在生理学和病理学中的作用，还因为它们在基因治疗、疫苗生产和诊断方面的新兴应用。与真核细胞产生的细胞外囊泡相比，细菌产生的细胞外囊泡鲜为人知，在革兰氏阴性菌中，主要类型被称为外膜囊泡（OMV）。在本研究中，我们首次展示了利用糖基磷脂酰肌醇（GPI）锚定蛋白对OMV进行功能性表面修饰，这一过程在真核细胞膜中被不同地描述为分子描绘或蛋白质工程，即GPI锚定的脂质部分插入细胞膜。通过将该过程转移到细菌囊泡上，我们可以在原核平台上生成完全具有真核蛋白质（在折叠和翻译后修饰方面）的杂种。我们能够证明两种不同的GPI蛋白可以展示在同一个OMV上。除了荧光标记蛋白外，细胞因子、生长因子和抗原也有可能被转移，从而为一种新型疫苗策略生成一个通用的模块化平台。
