Immobilization of recombinant vault nanoparticles on solid substrates.
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.
Xia
Yun
Y
School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798.
Ramgopal
Yamini
Y
Li
Hai
H
Shang
Lei
L
Srinivas
Parisa
P
Kickhoefer
Valerie A
VA
Rome
Leonard H
LH
Preiser
Peter R
PR
Boey
Freddy
F
Zhang
Hua
H
Venkatraman
Subbu S
SS
D000327:Adsorption; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D005898:Glass / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018625:Microscopy, Atomic Force; D053758:Nanoparticles / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry; D011437:Propylamines; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012821:Silanes / Q000737:chemistry; D055672:Static Electricity; D013499:Surface Properties; D020394:Vault Ribonucleoprotein Particles / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000099187:Major Vault Protein
D055767:Immobilized Proteins; D009842:Oligopeptides; D011437:Propylamines; D011994:Recombinant Proteins; D012821:Silanes; D020394:Vault Ribonucleoprotein Particles; D000099187:Major Vault Protein; C047981:arginyl-glycyl-aspartic acid; C477625:amino-propyl-triethoxysilane