Kurppa Katri, Hytönen Vesa P, Nakari-Setälä Tiina, Kulomaa Markku S, Linder Markus B
VTT Technical Research Institute of Finland, Tietotie 2, PO Box 1000, FI-02044 Espoo, Finland.
BioMediTech, University of Tampere and Tampere University Hospital, Biokatu 6, FI-33014 Tampere, Finland; Fimlab Laboratories, Biokatu 4, FI-33520 Tampere, Finland.
Colloids Surf B Biointerfaces. 2014 Aug 1;120:102-9. doi: 10.1016/j.colsurfb.2014.05.010. Epub 2014 May 22.
Control over the functionality of interfaces through biomolecular engineering is a central tool for nanoscale technology as well as many current applications of biology. In this work we designed fusion proteins that combined the surface adhesion and interfacial activity of a hydrophobin-protein together with the high affinity biotin-binding capability of an avidin-protein. We found that an overall architecture that was based on a circularly permuted version of avidin, dual-chain avidin, and hydrophobin gave a highly functional combination. The protein was produced in the filamentous fungus Trichoderma reesei and was efficiently purified using an aqueous two-phase partitioning procedure. The surface adhesive properties were widely different compared to wild-type avidin. Functional characterization showed that the protein assembled on hydrophobic surfaces as a thin layer even at very low concentrations and efficiently bound a biotinylated compound. The work shows how the challenge of creating a fusion protein with proteins that form multimers can be solved by structural design and how protein self-assembly can be used to efficiently functionalize interfaces.
通过生物分子工程控制界面功能是纳米技术以及当前许多生物学应用的核心工具。在这项工作中，我们设计了融合蛋白，它将疏水蛋白的表面粘附和界面活性与抗生物素蛋白的高亲和力生物素结合能力结合在一起。我们发现，基于抗生物素蛋白的循环置换版本、双链抗生物素蛋白和疏水蛋白的整体结构产生了高度功能性的组合。该蛋白在丝状真菌里氏木霉中产生，并使用双水相分配程序进行有效纯化。与野生型抗生物素蛋白相比，其表面粘附特性有很大差异。功能表征表明，该蛋白即使在非常低的浓度下也能在疏水表面组装成薄层，并有效结合生物素化化合物。这项工作展示了如何通过结构设计解决与形成多聚体的蛋白质创建融合蛋白的挑战，以及如何利用蛋白质自组装有效地使界面功能化。
