School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798.
ACS Nano. 2010 Mar 23;4(3):1417-24. doi: 10.1021/nn901167s.
Native vaults are nanoscale particles found abundantly in the cytoplasm of most eukaryotic cells. They have a capsule-like structure with a thin shell surrounding a "hollow" interior compartment. Recombinant vault particles were found to self-assemble following expression of the major vault protein (MVP) in a baculovirus expression system, and these particles are virtually identical to native vaults. Such particles have been recently studied as potential delivery vehicles. In this study, we focus on immobilization of vault particles on a solid substrate, such as glass, as a first step to study their interactions with cells. To this end, we first engineered the recombinant vaults by fusing two different tags to the C-terminus of MVP, a 3 amino acid RGD peptide and a 12 amino acid RGD-strep-tag peptide. We have demonstrated two strategies for immobilizing vaults on solid substrates. The barrel-and-cap structure of vault particles was observed for the first time, by atomic force microscopy (AFM), in a dry condition. This work proved the feasibility of immobilizing vault nanoparticles on a material surface, and the possibility of using vault nanoparticles as localized and sustainable drug carriers as well as a biocompatible surface moiety.
天然 vaults 是在大多数真核细胞细胞质中大量存在的纳米级颗粒。它们具有胶囊状结构，薄壳围绕着“中空”内部隔室。在杆状病毒表达系统中表达主要 vault 蛋白 (MVP) 后，发现重组 vault 颗粒会自组装，这些颗粒与天然 vaults 几乎完全相同。最近，这些颗粒已被作为潜在的递药载体进行研究。在这项研究中，我们专注于将 vault 颗粒固定在固体基质上，如玻璃上，作为研究它们与细胞相互作用的第一步。为此，我们首先通过将两个不同的标签融合到 MVP 的 C 末端，即 3 个氨基酸的 RGD 肽和 12 个氨基酸的 RGD-strep-tag 肽，对重组 vault 进行了工程改造。我们已经证明了两种将 vault 固定在固体基质上的策略。通过原子力显微镜 (AFM)，首次在干燥条件下观察到 vault 颗粒的桶帽结构。这项工作证明了将 vault 纳米颗粒固定在材料表面的可行性，以及将 vault 纳米颗粒用作局部和可持续的药物载体以及生物相容的表面部分的可能性。
