Schreiber Andreas, Stühn Lara G, Geissinger Süreyya E, Huber Matthias C, Schiller Stefan M
Center for Biological Systems Analysis, University of Freiburg; Faculty of Biology, University of Freiburg;
Center for Biological Systems Analysis, University of Freiburg; Faculty of Biology, University of Freiburg.
J Vis Exp. 2020 Apr 8(158). doi: 10.3791/60935.
Tailored proteinaceous building blocks are versatile candidates for the assembly of supramolecular structures such as minimal cells, drug delivery vehicles and enzyme scaffolds. Due to their biocompatibility and tunability on the genetic level, Elastin-like proteins (ELP) are ideal building blocks for biotechnological and biomedical applications. Nevertheless, the assembly of protein based supramolecular structures with distinct physiochemical properties and good encapsulation potential remains challenging. Here we provide two efficient protocols for guided self-assembly of amphiphilic ELPs into supramolecular protein architectures such as spherical coacervates, fibers and stable vesicles. The presented assembly protocols generate Protein Membrane-Based Compartments (PMBCs) based on ELPs with adaptable physicochemical properties. PMBCs demonstrate phase separation behavior and reveal method dependent membrane fusion and are able to encapsulate chemically diverse fluorescent cargo molecules. The resulting PMBCs have a high application potential as a drug formulation and delivery platform, artificial cell, and compartmentalized reaction space.
定制的蛋白质构建模块是用于组装超分子结构（如最小细胞、药物递送载体和酶支架）的通用候选物。由于其生物相容性和在基因水平上的可调节性，类弹性蛋白（ELP）是生物技术和生物医学应用的理想构建模块。然而，组装具有独特物理化学性质和良好包封潜力的基于蛋白质的超分子结构仍然具有挑战性。在这里，我们提供了两种有效的方案，用于将两亲性ELP引导自组装成超分子蛋白质结构，如球形凝聚物、纤维和稳定的囊泡。所提出的组装方案基于具有可适应物理化学性质的ELP生成基于蛋白质膜的隔室（PMBC）。PMBC表现出相分离行为，揭示了方法依赖性的膜融合，并且能够封装化学性质多样的荧光货物分子。所得的PMBC作为药物制剂和递送平台、人工细胞和分隔反应空间具有很高的应用潜力。
