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Structural characterization of site-modified nanocapsid with monodispersed gold clusters

Hepatitis E Virus-like particles self-assemble in to noninfectious nanocapsids that are resistant to proteolytic/acidic mucosal delivery conditions. Previously, the nanocapsid was engineered to specifically bind and enter breast cancer cells, where successful tumor targeting was demonstrated in anim...

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Autores principales: Stark, Marie C, Baikoghli, Mo A, Lahtinen, Tanja, Malola, Sami, Xing, Li, Nguyen, Michelle, Nguyen, Marina, Sikaroudi, Aria, Marjomäki, Varpu, Häkkinen, Hannu, Cheng, R Holland
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5719084/
https://www.ncbi.nlm.nih.gov/pubmed/29213060
http://dx.doi.org/10.1038/s41598-017-17171-x
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author Stark, Marie C
Baikoghli, Mo A
Lahtinen, Tanja
Malola, Sami
Xing, Li
Nguyen, Michelle
Nguyen, Marina
Sikaroudi, Aria
Marjomäki, Varpu
Häkkinen, Hannu
Cheng, R Holland
author_facet Stark, Marie C
Baikoghli, Mo A
Lahtinen, Tanja
Malola, Sami
Xing, Li
Nguyen, Michelle
Nguyen, Marina
Sikaroudi, Aria
Marjomäki, Varpu
Häkkinen, Hannu
Cheng, R Holland
author_sort Stark, Marie C
collection PubMed
description Hepatitis E Virus-like particles self-assemble in to noninfectious nanocapsids that are resistant to proteolytic/acidic mucosal delivery conditions. Previously, the nanocapsid was engineered to specifically bind and enter breast cancer cells, where successful tumor targeting was demonstrated in animal models. In the present study, the nanocapsid surface was modified with a solvent-exposed cysteine to conjugate monolayer protected gold nanoclusters (AuNC). Unlike commercially available gold nanoparticles, AuNCs monodisperse in water and are composed of a discrete number of gold atoms, forming a crystalline gold core. Au(102) pMBA(44) (Au(102)) was an ideal conjugate given its small 2.5 nm size and detectability in cryoEM. Au(102) was bound directly to nanocapsid surface cysteines via direct ligand exchange. In addition, Au(102) was functionalized with a maleimide linker (Au(102)_C(6)MI) for maleimide-thiol conjugation to nanocapsid cysteines. The AuNC-bound nanocapsid constructs were conjugated in various conditions. We found Au(102)_C(6)MI to bind nanocapsid more efficiently, while Au(102) remained more soluble over time. Nanocapsids conjugated to Au(102)_C(6)MI were imaged in cryoEM for single particle reconstruction to localize AuNC position on the nanocapsid surface. We resolved five unique high intensity volumes that formed a ring-shaped density at the 5-fold symmetry center. This finding was further supported by independent rigid modeling.
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spelling pubmed-57190842017-12-08 Structural characterization of site-modified nanocapsid with monodispersed gold clusters Stark, Marie C Baikoghli, Mo A Lahtinen, Tanja Malola, Sami Xing, Li Nguyen, Michelle Nguyen, Marina Sikaroudi, Aria Marjomäki, Varpu Häkkinen, Hannu Cheng, R Holland Sci Rep Article Hepatitis E Virus-like particles self-assemble in to noninfectious nanocapsids that are resistant to proteolytic/acidic mucosal delivery conditions. Previously, the nanocapsid was engineered to specifically bind and enter breast cancer cells, where successful tumor targeting was demonstrated in animal models. In the present study, the nanocapsid surface was modified with a solvent-exposed cysteine to conjugate monolayer protected gold nanoclusters (AuNC). Unlike commercially available gold nanoparticles, AuNCs monodisperse in water and are composed of a discrete number of gold atoms, forming a crystalline gold core. Au(102) pMBA(44) (Au(102)) was an ideal conjugate given its small 2.5 nm size and detectability in cryoEM. Au(102) was bound directly to nanocapsid surface cysteines via direct ligand exchange. In addition, Au(102) was functionalized with a maleimide linker (Au(102)_C(6)MI) for maleimide-thiol conjugation to nanocapsid cysteines. The AuNC-bound nanocapsid constructs were conjugated in various conditions. We found Au(102)_C(6)MI to bind nanocapsid more efficiently, while Au(102) remained more soluble over time. Nanocapsids conjugated to Au(102)_C(6)MI were imaged in cryoEM for single particle reconstruction to localize AuNC position on the nanocapsid surface. We resolved five unique high intensity volumes that formed a ring-shaped density at the 5-fold symmetry center. This finding was further supported by independent rigid modeling. Nature Publishing Group UK 2017-12-06 /pmc/articles/PMC5719084/ /pubmed/29213060 http://dx.doi.org/10.1038/s41598-017-17171-x Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Stark, Marie C
Baikoghli, Mo A
Lahtinen, Tanja
Malola, Sami
Xing, Li
Nguyen, Michelle
Nguyen, Marina
Sikaroudi, Aria
Marjomäki, Varpu
Häkkinen, Hannu
Cheng, R Holland
Structural characterization of site-modified nanocapsid with monodispersed gold clusters
title Structural characterization of site-modified nanocapsid with monodispersed gold clusters
title_full Structural characterization of site-modified nanocapsid with monodispersed gold clusters
title_fullStr Structural characterization of site-modified nanocapsid with monodispersed gold clusters
title_full_unstemmed Structural characterization of site-modified nanocapsid with monodispersed gold clusters
title_short Structural characterization of site-modified nanocapsid with monodispersed gold clusters
title_sort structural characterization of site-modified nanocapsid with monodispersed gold clusters
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5719084/
https://www.ncbi.nlm.nih.gov/pubmed/29213060
http://dx.doi.org/10.1038/s41598-017-17171-x
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