将细菌作为超分子自组装单分子层中的活性组分通过动态纳米尺度相互作用进行整合
Incorporating Bacteria as a Living Component in Supramolecular Self-Assembled Monolayers through Dynamic Nanoscale Interactions
作者:Shrikrishnan Sankaran;Mustafa Can Kiren;Pascal Jonkheijm;
关键词:supramolecular,Cucurbit[8]uril,E. coli,bacterial display,eCPX,knottins,aggregation,self-assembled monolayers
DOI:https://doi.org/10.1021/acsnano.5b00694
发表时间:2015年
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摘要
超分子组装体通过非共价相互作用形成,已经变得特别吸引人,用来开发动态和响应型体系以应对纳米尺度的生物系统。南瓜[8]葫芦[8](CB[8]),一种南瓜形状的大环宿主分子,已成功用于构建各种自组装体系,用于生物医学应用,因为它能同时在其腔内结合两个芳香族客体分子。这种结构也可以设计以对外部刺激作出响应。将生物体纳入这种超分子结构作为主动成分会为这些体系的功能增添新的维度。为了实现这一点,我们通过基因修饰转膜蛋白,在细菌表面上引入了超分子功能,以展示CB[8]的结合基序作为胱氨酸稳定的小蛋白的一部分。我们成功证实了细菌表面的超分子结构单元能够特异性结合CB[8]并形成多个细胞间的三元复合物,导致细菌溶液的聚集。我们进行了各种聚集实验,以了解CB[8]如何与这种细菌菌株相互作用,并且证明它可以通过竞争配体进行化学逆转。为了确认这个菌株能够与基于CB[8]的结构体系结合,我们展示了这种细菌能够粘附在金材料上的CB[8]自组装单分子膜,并且仍然在数小时内保持相当的运动能力,表明这个系统潜在地可以用于开发超分子细菌生物马达。这种细菌菌株还可以与其他基于CB[8]的体系结合,比如纳米粒子、囊泡和水凝胶。
Abstract
Supramolecular assemblies, formed through noncovalent interactions, has become particularly attractive to develop dynamic and responsive architectures to address living systems at the nanoscale. Cucurbit[8]uril (CB[8]), a pumpkin shaped macrocylic host molecule, has been successfully used to construct various self-assembled architectures for biomedical applications since it can simultaneously bind two aromatic guest molecules within its cavity. Such architectures can also be designed to respond to external stimuli. Integrating living organisms as an active component into such supramolecular architectures would add a new dimension to the capabilities of such systems. To achieve this, we have incorporated supramolecular functionality at the bacterial surface by genetically modifying a transmembrane protein to display a CB[8]-binding motif as part of a cystine-stabilized miniprotein. We were able to confirm that this supramolecular motif on the bacterial surface specifically binds CB[8] and forms multiple intercellular ternary complexes leading to aggregation of the bacterial solution. We performed various aggregation experiments to understand how CB[8] interacts with this bacterial strain and also demonstrate that it can be chemically reversed using a competitor. To confirm that this strain can be incorporated with a CB[8] based architecture, we show that the bacterial cells were able to adhere to CB[8] self-assembled monolayers (SAMs) on gold and still retain considerable motility for several hours, indicating that the system can potentially be used to develop supramolecular bacterial biomotors. The bacterial strain also has the potential to be combined with other CB[8] based architectures like nanoparticles, vesicles and hydrogels.
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