View : 1 Download: 0
Heparin Binding to an Engineered Virus-like Nanoparticle Antagonist
- Heparin Binding to an Engineered Virus-like Nanoparticle Antagonist
- Gale A.J.; Udit A.K.; Cheong H.Y.; Groner M.; Hong K.; Lynch B.; Hollingsworth W.R.; Polonskaya Z.; Rhee J.-K.; Baksh M.M.; Finn M.G.
- Ewha Authors
- SCOPUS Author ID
- Issue Date
- Journal Title
- vol. 18, no. 12, pp. 4113 - 4120
- American Chemical Society
- SCI; SCIE; SCOPUS
- The anticoagulant activity of heparin administered during medical interventions must be reversed to restore normal clotting, typically by titrating with protamine. Given the acute toxicity associated with protamine, we endeavored to generate safer heparin antagonists by engineering bacteriophage Qβ virus-like particles (VLPs) to display motifs that bind heparin. A particle bearing a single amino acid change from wild-type (T18R) was identified as a promising candidate for heparin antagonism. Surface potential maps generated through molecular modeling reveal that the T18R mutation adds synergistically to adjacent positive charges on the particle surface, resulting in a large solvent-accessible cationic region that is replicated 180 times over the capsid. Chromatography using a heparin-sepharose column confirmed a strong interaction between heparin and the T18R particle. Binding studies using fluorescein-labeled heparin (HepFL) resulted in a concentration-dependent change in fluorescence intensity, which could be perturbed by the addition of unlabeled heparin. Analysis of the fluorescence data yielded a dissociation constant of approximately 1 nM and a 1:1 binding stoichiometry for HepFL:VLP. Dynamic light scattering (DLS) experiments suggested that T18R forms discrete complexes with heparin when the VLP:heparin molar ratios are equivalent, and in vitro clotting assays confirmed the 1:1 binding stoichiometry as full antagonism of heparin is achieved. Biolayer interferometry and backscattering interferometry corroborated the strong interaction of T18R with heparin, yielding Kd ∼ 1-10 nM. These biophysical measurements further validated T18R, and VLPs in general, for potential clinical use as effective, nontoxic heparin antagonists. © 2017 American Chemical Society.
- Appears in Collections:
- 엘텍공과대학 > 식품공학전공 > Journal papers
- Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.