Gagic Dragana, Ciric Milica, Wen Wesley X, Ng Filomena, Rakonjac Jasna
Institute of Fundamental Sciences, Massey UniversityPalmerston North, New Zealand; Animal Science, Grasslands Research Centre, AgResearch Ltd, Palmerston NorthNew Zealand.
Institute of Fundamental Sciences, Massey University Palmerston North, New Zealand.
Front Microbiol. 2016 Apr 7;7:429. doi: 10.3389/fmicb.2016.00429. eCollection 2016.
Microbial surface and secreted proteins (the secretome) contain a large number of proteins that interact with other microbes, host and/or environment. These proteins are exported by the coordinated activities of the protein secretion machinery present in the cell. A group of bacteriophage, called filamentous phage, have the ability to hijack bacterial protein secretion machinery in order to amplify and assemble via a secretion-like process. This ability has been harnessed in the use of filamentous phage of Escherichia coli in biotechnology applications, including screening large libraries of variants for binding to "bait" of interest, from tissues in vivo to pure proteins or even inorganic substrates. In this review we discuss the roles of secretome proteins in pathogenic and non-pathogenic bacteria and corresponding secretion pathways. We describe the basics of phage display technology and its variants applied to discovery of bacterial proteins that are implicated in colonization of host tissues and pathogenesis, as well as vaccine candidates through filamentous phage display library screening. Secretome selection aided by next-generation sequence analysis was successfully applied for selective display of the secretome at a microbial community scale, the latter revealing the richness of secretome functions of interest and surprising versatility in filamentous phage display of secretome proteins from large number of Gram-negative as well as Gram-positive bacteria and archaea.
微生物表面蛋白和分泌蛋白（分泌组）包含大量与其他微生物、宿主和/或环境相互作用的蛋白质。这些蛋白质通过细胞中存在的蛋白质分泌机制的协同活动被输出。一类被称为丝状噬菌体的噬菌体，能够劫持细菌蛋白质分泌机制，以便通过类似分泌的过程进行扩增和组装。这种能力已被用于生物技术应用中，利用大肠杆菌丝状噬菌体筛选大量变体文库，以寻找与感兴趣的“诱饵”结合的物质，从体内组织到纯蛋白质甚至无机底物。在这篇综述中，我们讨论了分泌组蛋白在致病性和非致病性细菌中的作用以及相应的分泌途径。我们描述了噬菌体展示技术及其变体的基本原理，这些技术应用于发现与宿主组织定植和发病机制相关的细菌蛋白，以及通过丝状噬菌体展示文库筛选寻找候选疫苗。借助下一代序列分析的分泌组选择已成功应用于在微生物群落规模上选择性展示分泌组，后者揭示了感兴趣的分泌组功能的丰富性以及丝状噬菌体展示来自大量革兰氏阴性菌、革兰氏阳性菌和古菌的分泌组蛋白时令人惊讶的多功能性。
