Daudey Geert A, Shen Mengjie, Singhal Ankush, van der Est Patrick, Sevink G J Agur, Boyle Aimee L, Kros Alexander
Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University P.O. Box 9502 2300 RA Leiden The Netherlands
Macromolecular Biochemistry, Leiden Institute of Chemistry, Leiden University P.O. Box 9502 2300 RA Leiden The Netherlands.
Chem Sci. 2021 Sep 22;12(41):13782-13792. doi: 10.1039/d0sc06635d. eCollection 2021 Oct 27.
Biological membrane fusion is a highly specific and coordinated process as a multitude of vesicular fusion events proceed simultaneously in a complex environment with minimal off-target delivery. In this study, we develop a liposomal fusion model system with specific recognition using lipidated derivatives of a set of four designed heterodimeric coiled coil (CC) peptide pairs. Content mixing was only obtained between liposomes functionalized with complementary peptides, demonstrating both fusogenic activity of CC peptides and the specificity of this model system. The diverse peptide fusogens revealed important relationships between the fusogenic efficacy and the peptide characteristics. The fusion efficiency increased from 20% to 70% as affinity between complementary peptides decreased, (from ≈ 10 to 10 M), and fusion efficiency also increased due to more pronounced asymmetric role-playing of membrane interacting 'K' peptides and homodimer-forming 'E' peptides. Furthermore, a new and highly fusogenic CC pair (E/P1) was discovered, providing an orthogonal peptide triad with the fusogenic CC pairs P2/P2 and P3/P3. This E/P1 pair was revealed, molecular dynamics simulations, to have a shifted heptad repeat that can accommodate mismatched asparagine residues. These results will have broad implications not only for the fundamental understanding of CC design and how asparagine residues can be accommodated within the hydrophobic core, but also for drug delivery systems by revealing the necessary interplay of efficient peptide fusogens and enabling the targeted delivery of different carrier vesicles at various peptide-functionalized locations.
生物膜融合是一个高度特异且协调的过程，因为众多囊泡融合事件在复杂环境中同时进行，且脱靶递送极少。在本研究中，我们利用一组四个设计的异源二聚卷曲螺旋（CC）肽对的脂化衍生物开发了一种具有特异性识别的脂质体融合模型系统。仅在用互补肽功能化的脂质体之间观察到内容物混合，这既证明了CC肽的融合活性，也证明了该模型系统的特异性。多种肽融合剂揭示了融合效力与肽特性之间的重要关系。随着互补肽之间的亲和力降低（从≈10到10 M），融合效率从20%提高到70%，并且由于膜相互作用的“K”肽和形成同二聚体的“E”肽更明显的不对称作用，融合效率也有所提高。此外，还发现了一种新的高融合性CC对（E/P1），它与融合性CC对P2/P2和P3/P3形成了一个正交肽三联体。通过分子动力学模拟发现，该E/P1对具有移位的七肽重复序列，能够容纳错配的天冬酰胺残基。这些结果不仅对CC设计的基本理解以及天冬酰胺残基如何能容纳在疏水核心内具有广泛影响，而且通过揭示高效肽融合剂的必要相互作用以及实现不同载体囊泡在各种肽功能化位置的靶向递送，对药物递送系统也具有广泛影响。
