Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles
The folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engine...
Main Authors: | , , , , , , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2018-05-01
|
Series: | Frontiers in Immunology |
Subjects: | |
Online Access: | http://journal.frontiersin.org/article/10.3389/fimmu.2018.01093/full |
id |
doaj-7d5a9fc141994285863439d81f58903e |
---|---|
record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Young-Seok Kim Young-Seok Kim Ahyun Son Jihoon Kim Jihoon Kim Soon Bin Kwon Soon Bin Kwon Myung Hee Kim Paul Kim Paul Kim Jieun Kim Young Ho Byun Jemin Sung Jemin Sung Jinhee Lee Jinhee Lee Ji Eun Yu Ji Eun Yu Chan Park Chan Park Yeon-Sook Kim Nam-Hyuk Cho Nam-Hyuk Cho Jun Chang Baik L. Seong Baik L. Seong |
spellingShingle |
Young-Seok Kim Young-Seok Kim Ahyun Son Jihoon Kim Jihoon Kim Soon Bin Kwon Soon Bin Kwon Myung Hee Kim Paul Kim Paul Kim Jieun Kim Young Ho Byun Jemin Sung Jemin Sung Jinhee Lee Jinhee Lee Ji Eun Yu Ji Eun Yu Chan Park Chan Park Yeon-Sook Kim Nam-Hyuk Cho Nam-Hyuk Cho Jun Chang Baik L. Seong Baik L. Seong Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles Frontiers in Immunology nanoparticle virus-like particle chaperone ferritin Middle East respiratory syndrome coronavirus receptor-binding domain |
author_facet |
Young-Seok Kim Young-Seok Kim Ahyun Son Jihoon Kim Jihoon Kim Soon Bin Kwon Soon Bin Kwon Myung Hee Kim Paul Kim Paul Kim Jieun Kim Young Ho Byun Jemin Sung Jemin Sung Jinhee Lee Jinhee Lee Ji Eun Yu Ji Eun Yu Chan Park Chan Park Yeon-Sook Kim Nam-Hyuk Cho Nam-Hyuk Cho Jun Chang Baik L. Seong Baik L. Seong |
author_sort |
Young-Seok Kim |
title |
Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles |
title_short |
Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles |
title_full |
Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles |
title_fullStr |
Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles |
title_full_unstemmed |
Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus Nanoparticles |
title_sort |
chaperna-mediated assembly of ferritin-based middle east respiratory syndrome-coronavirus nanoparticles |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2018-05-01 |
description |
The folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engineered NPs are refractory to soluble expression and fail to assemble as designed, presenting major challenges in the manufacturing process. The failure is due to a lack of understanding of the kinetic pathways and enabling technical platforms to ensure successful folding of the monomer antigens into regular assemblages. Capitalizing on a novel function of RNA as a molecular chaperone (chaperna: chaperone + RNA), we provide a robust protein-folding vehicle that may be implemented to NP assembly in bacterial hosts. The receptor-binding domain (RBD) of Middle East respiratory syndrome-coronavirus (MERS-CoV) was fused with the RNA-interaction domain (RID) and bacterioferritin, and expressed in Escherichia coli in a soluble form. Site-specific proteolytic removal of the RID prompted the assemblage of monomers into NPs, which was confirmed by electron microscopy and dynamic light scattering. The mutations that affected the RNA binding to RBD significantly increased the soluble aggregation into amorphous structures, reducing the overall yield of NPs of a defined size. This underscored the RNA-antigen interactions during NP assembly. The sera after mouse immunization effectively interfered with the binding of MERS-CoV RBD to the cellular receptor hDPP4. The results suggest that RNA-binding controls the overall kinetic network of the antigen folding pathway in favor of enhanced assemblage of NPs into highly regular and immunologically relevant conformations. The concentration of the ion Fe2+, salt, and fusion linker also contributed to the assembly in vitro, and the stability of the NPs. The kinetic “pace-keeping” role of chaperna in the super molecular assembly of antigen monomers holds promise for the development and delivery of NPs and virus-like particles as recombinant vaccines and for serological detection of viral infections. |
topic |
nanoparticle virus-like particle chaperone ferritin Middle East respiratory syndrome coronavirus receptor-binding domain |
url |
http://journal.frontiersin.org/article/10.3389/fimmu.2018.01093/full |
work_keys_str_mv |
AT youngseokkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT youngseokkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT ahyunson chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jihoonkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jihoonkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT soonbinkwon chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT soonbinkwon chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT myungheekim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT paulkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT paulkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jieunkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT younghobyun chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jeminsung chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jeminsung chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jinheelee chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jinheelee chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jieunyu chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT jieunyu chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT chanpark chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT chanpark chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT yeonsookkim chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT namhyukcho chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT namhyukcho chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT junchang chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT baiklseong chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles AT baiklseong chapernamediatedassemblyofferritinbasedmiddleeastrespiratorysyndromecoronavirusnanoparticles |
_version_ |
1725293866342416384 |
spelling |
doaj-7d5a9fc141994285863439d81f58903e2020-11-25T00:39:18ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-05-01910.3389/fimmu.2018.01093346199Chaperna-Mediated Assembly of Ferritin-Based Middle East Respiratory Syndrome-Coronavirus NanoparticlesYoung-Seok Kim0Young-Seok Kim1Ahyun Son2Jihoon Kim3Jihoon Kim4Soon Bin Kwon5Soon Bin Kwon6Myung Hee Kim7Paul Kim8Paul Kim9Jieun Kim10Young Ho Byun11Jemin Sung12Jemin Sung13Jinhee Lee14Jinhee Lee15Ji Eun Yu16Ji Eun Yu17Chan Park18Chan Park19Yeon-Sook Kim20Nam-Hyuk Cho21Nam-Hyuk Cho22Jun Chang23Baik L. Seong24Baik L. Seong25Department of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaGraduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaLife Science and Biotechnology, Underwood International College, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaDivision of Infectious Diseases, Department of Internal Medicine, Chungnam National University School of Medicine, Daejeon, South KoreaDepartment of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, South KoreaDepartment of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South KoreaGraduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, South KoreaDepartment of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul, South KoreaVaccine Translational Research Center, Yonsei University, Seoul, South KoreaThe folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engineered NPs are refractory to soluble expression and fail to assemble as designed, presenting major challenges in the manufacturing process. The failure is due to a lack of understanding of the kinetic pathways and enabling technical platforms to ensure successful folding of the monomer antigens into regular assemblages. Capitalizing on a novel function of RNA as a molecular chaperone (chaperna: chaperone + RNA), we provide a robust protein-folding vehicle that may be implemented to NP assembly in bacterial hosts. The receptor-binding domain (RBD) of Middle East respiratory syndrome-coronavirus (MERS-CoV) was fused with the RNA-interaction domain (RID) and bacterioferritin, and expressed in Escherichia coli in a soluble form. Site-specific proteolytic removal of the RID prompted the assemblage of monomers into NPs, which was confirmed by electron microscopy and dynamic light scattering. The mutations that affected the RNA binding to RBD significantly increased the soluble aggregation into amorphous structures, reducing the overall yield of NPs of a defined size. This underscored the RNA-antigen interactions during NP assembly. The sera after mouse immunization effectively interfered with the binding of MERS-CoV RBD to the cellular receptor hDPP4. The results suggest that RNA-binding controls the overall kinetic network of the antigen folding pathway in favor of enhanced assemblage of NPs into highly regular and immunologically relevant conformations. The concentration of the ion Fe2+, salt, and fusion linker also contributed to the assembly in vitro, and the stability of the NPs. The kinetic “pace-keeping” role of chaperna in the super molecular assembly of antigen monomers holds promise for the development and delivery of NPs and virus-like particles as recombinant vaccines and for serological detection of viral infections.http://journal.frontiersin.org/article/10.3389/fimmu.2018.01093/fullnanoparticlevirus-like particlechaperoneferritinMiddle East respiratory syndrome coronavirusreceptor-binding domain |