Haloferax volcanii microproteins that contain two CPXCG motifs indicative for one-domain zinc finger proteins

该项目在优先计划SPP2002“原核生物中的小蛋白”的框架内，集中在嗜盐古生物Haloferax Volcanii的非常小的蛋白上，这些蛋白包含两个C(P)XCG基序，因此是一个结构域的锌指蛋白。在第一次授予期间，16个缺失突变体的表型特征揭示了嗜盐古菌C(P)XCG蛋白具有很高的生物学意义。一种名为HVO_2753的蛋白质已经被详细鉴定，并解决了它的核磁共振溶液结构。在第二个授权期，将产生一个进一步的C(P)XCG蛋白质基因的缺失突变集合并对其进行鉴定。将建立一种新的“隐藏的硫醇蛋白质组学”方法，并用于不同环境条件下小锌指蛋白的差异生产的并行表征。我们还将进一步详细研究HVO_2753和其他几个具有高度生物学意义的C(P)XCG蛋白，如揭示选定的单一氨基酸在体内对蛋白质功能的重要性，确定相互作用伙伴，并进行进一步的结构研究。综上所述，该项目将导致对小锌指蛋白在海洋生物中的生物学作用和分子机制的前所未有的深入了解。

This project in the framework of the Priority Program SPP2002 “Small proteins in prokaryotes” concentrates on very small proteins of the halophilic archaeaon Haloferax volcanii, which contain two C(P)XCG motifs, and, thus, are one-domain zinc finger proteins. In the first granting period, phenotypic characterization of 16 deletion mutants has revealed the high biological significance of haloarchaeal C(P)XCG proteins. One protein, HVO_2753, has been characterized in detail, and its NMR solution structure was solved.In the second granting period, a deletion mutant collection of further C(P)XCG protein genes will be generated and characterized. A novel approach of “hidden thiol proteomics” will be established and used for the parallel characterization of differential production of small zinc finger proteins under various environmental conditions. HVO_2753 and a few additional C(P)XCG proteins of high biological significance will be further characterized in detail, e.g. the importance of selected single amino acids for protein function in vivo will be unraveled, interaction partners will be identified, and further structural studies will be performed. In summary, the project will lead to an unprecedented insight into the biological roles and molecular mechanisms of small zinc finger proteins in haloarchaea.