Kuan Seah Ling, Förtsch Christina, Ng David Yuen Wah, Fischer Stephan, Tokura Yu, Liu Weina, Wu Yuzhou, Koynov Kaloian, Barth Holger, Weil Tanja
Institute of Organic Chemistry III - Macromolecular Chemistry & Biomaterials, University of Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany.
Institute of Pharmacology and Toxicology, University of Ulm Medical Center, Albert-Einstein-Allee 11, 89081, Ulm, Germany.
Macromol Biosci. 2016 Jun;16(6):803-10. doi: 10.1002/mabi.201500417. Epub 2016 Feb 2.
Nature has provided a highly optimized toolbox in bacterial endotoxins with precise functions dictated by their clear structural division. Inspired by this streamlined design, a supramolecular approach capitalizing on the strong biomolecular (streptavidin (SA))-biotin interactions is reported herein to prepare two multipartite fusion constructs, which involves the generation 2.0 (D2) or generation 3.0 (D3) polyamidoamine-dendronized transporter proteins (dendronized streptavidin (D3SA) and dendronized human serum albumin (D2HSA)) non-covalently fused to the C3bot1 enzyme from Clostridium botulinum, a potent and specific Rho-inhibitor. The fusion constructs, D3SA-C3 and D2HSA-C3, represent the first examples of dendronized protein transporters that are fused to the C3 enzyme, and it is successfully demonstrated that the C3 Rho-inhibitor is delivered into the cytosol of mammalian cells as determined from the characteristic C3-mediated changes in cell morphology and confocal microscopy. The design circumvents the low uptake of the C3 enzyme by eukaryotic cells and holds great promise for reprogramming the properties of toxin enzymes using a supramolecular approach to broaden their therapeutic applications.
自然界在细菌内毒素中提供了一个高度优化的工具箱，其精确的功能由清晰的结构划分决定。受这种简化设计的启发，本文报道了一种利用强大的生物分子（链霉亲和素（SA））-生物素相互作用的超分子方法，以制备两种多部分融合构建体，其中涉及非共价融合到来自肉毒杆菌的C3bot1酶（一种有效且特异性的Rho抑制剂）的第2代（D2）或第3代（D3）聚酰胺胺树枝状化转运蛋白（树枝状化链霉亲和素（D3SA）和树枝状化人血清白蛋白（D2HSA））。融合构建体D3SA-C3和D2HSA-C3代表了与C3酶融合的树枝状化蛋白转运体的首个实例，并且成功证明，根据细胞形态和共聚焦显微镜下C3介导的特征性变化确定，C3 Rho抑制剂被递送至哺乳动物细胞的细胞质中。该设计规避了真核细胞对C3酶的低摄取，并有望通过超分子方法重新编程毒素酶的特性，以拓宽其治疗应用。
