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Title: Nanoparticle-assisted tubulin assembly is environment dependent
Nanomaterials acquire a biomolecular corona upon introduction to biological media, leading to biological transformations such as changes in protein function, unmasking of epitopes, and protein fibrilization. Ex vivo studies to investigate the effect of nanoparticles on protein–protein interactions are typically performed in buffer and are rarely measured quantitatively in live cells. Here, we measure the differential effect of silica nanoparticles on protein association in vitro vs. in mammalian cells. BtubA and BtubB are a pair of bacterial tubulin proteins identified inProsthecobacterstrains that self-assemble like eukaryotic tubulin, first into dimers and then into microtubules in vitro or in vivo. Förster resonance energy transfer labeling of each of the Btub monomers with a donor (mEGFP) and acceptor (mRuby3) fluorescent protein provides a quantitative tool to measure their binding interactions in the presence of unfunctionalized silica nanoparticles in buffer and in cells using fluorescence spectroscopy and microscopy. We show that silica nanoparticles enhance BtubAB dimerization in buffer due to protein corona formation. However, these nanoparticles have little effect on bacterial tubulin self-assembly in the complex mammalian cellular environment. Thus, the effect of nanomaterials on protein–protein interactions may not be readily translated from the test tube to the cell in the absence of particle surface functionalization that can enable targeted protein–nanoparticle interactions to withstand competitive binding in the nanoparticle corona from other biomolecules.  more » « less
Award ID(s):
2001611
PAR ID:
10598016
Author(s) / Creator(s):
; ; ;
Publisher / Repository:
Proceedings of the National Academy of Sciences USA
Date Published:
Journal Name:
Proceedings of the National Academy of Sciences
Volume:
121
Issue:
28
ISSN:
0027-8424
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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