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Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin

We have recently reported that calreticulin (CRT), a luminal resident protein, can be found in the sera of patients with rheumatoid arthritis and also that recombinant CRT (rCRT) exhibits extraordinarily strong immunological activities. We herein further demonstrate that rCRT fragments 18–412 (rCRT/...

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Autores principales: Huang, Shang-Hui, Zhao, Li-Xiang, Hong, Chao, Duo, Cui-Cui, Guo, Bing-Nan, Zhang, Li-Juan, Gong, Zheng, Xiong, Si-Dong, Gong, Fang-Yuan, Gao, Xiao-Ming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3677884/
https://www.ncbi.nlm.nih.gov/pubmed/23762269
http://dx.doi.org/10.1371/journal.pone.0064951
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author Huang, Shang-Hui
Zhao, Li-Xiang
Hong, Chao
Duo, Cui-Cui
Guo, Bing-Nan
Zhang, Li-Juan
Gong, Zheng
Xiong, Si-Dong
Gong, Fang-Yuan
Gao, Xiao-Ming
author_facet Huang, Shang-Hui
Zhao, Li-Xiang
Hong, Chao
Duo, Cui-Cui
Guo, Bing-Nan
Zhang, Li-Juan
Gong, Zheng
Xiong, Si-Dong
Gong, Fang-Yuan
Gao, Xiao-Ming
author_sort Huang, Shang-Hui
collection PubMed
description We have recently reported that calreticulin (CRT), a luminal resident protein, can be found in the sera of patients with rheumatoid arthritis and also that recombinant CRT (rCRT) exhibits extraordinarily strong immunological activities. We herein further demonstrate that rCRT fragments 18–412 (rCRT/18-412), rCRT/39-272, rCRT/120-308 and rCRT/120-250 can self-oligomerize in solution and are 50–100 fold more potent than native CRT (nCRT, isolated from mouse livers) in activating macrophages in vitro. We narrowed down the active site of CRT to residues 150–230, the activity of which also depends on dimerization. By contrast, rCRT/18-197 is almost completely inactive. When rCRT/18-412 is fractionated into oligomers and monomers by gel filtration, the oligomers maintain most of their immunological activities in terms of activating macrophages in vitro and inducing specific antibodies in vivo, while the monomers were much less active by comparison. Additionally, rCRT/18-412 oligomers are much better than monomers in binding to, and uptake by, macrophages. Inhibition of macrophage endocytosis partially blocks the stimulatory effect of rCRT/18-412. We conclude that the immunologically active site of CRT maps between residues 198–230 and that soluble CRT could acquire potent immuno-pathological activities in microenvironments favoring its oligomerization.
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spelling pubmed-36778842013-06-12 Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin Huang, Shang-Hui Zhao, Li-Xiang Hong, Chao Duo, Cui-Cui Guo, Bing-Nan Zhang, Li-Juan Gong, Zheng Xiong, Si-Dong Gong, Fang-Yuan Gao, Xiao-Ming PLoS One Research Article We have recently reported that calreticulin (CRT), a luminal resident protein, can be found in the sera of patients with rheumatoid arthritis and also that recombinant CRT (rCRT) exhibits extraordinarily strong immunological activities. We herein further demonstrate that rCRT fragments 18–412 (rCRT/18-412), rCRT/39-272, rCRT/120-308 and rCRT/120-250 can self-oligomerize in solution and are 50–100 fold more potent than native CRT (nCRT, isolated from mouse livers) in activating macrophages in vitro. We narrowed down the active site of CRT to residues 150–230, the activity of which also depends on dimerization. By contrast, rCRT/18-197 is almost completely inactive. When rCRT/18-412 is fractionated into oligomers and monomers by gel filtration, the oligomers maintain most of their immunological activities in terms of activating macrophages in vitro and inducing specific antibodies in vivo, while the monomers were much less active by comparison. Additionally, rCRT/18-412 oligomers are much better than monomers in binding to, and uptake by, macrophages. Inhibition of macrophage endocytosis partially blocks the stimulatory effect of rCRT/18-412. We conclude that the immunologically active site of CRT maps between residues 198–230 and that soluble CRT could acquire potent immuno-pathological activities in microenvironments favoring its oligomerization. Public Library of Science 2013-06-10 /pmc/articles/PMC3677884/ /pubmed/23762269 http://dx.doi.org/10.1371/journal.pone.0064951 Text en © 2013 Huang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Huang, Shang-Hui
Zhao, Li-Xiang
Hong, Chao
Duo, Cui-Cui
Guo, Bing-Nan
Zhang, Li-Juan
Gong, Zheng
Xiong, Si-Dong
Gong, Fang-Yuan
Gao, Xiao-Ming
Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin
title Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin
title_full Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin
title_fullStr Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin
title_full_unstemmed Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin
title_short Self-Oligomerization Is Essential for Enhanced Immunological Activities of Soluble Recombinant Calreticulin
title_sort self-oligomerization is essential for enhanced immunological activities of soluble recombinant calreticulin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3677884/
https://www.ncbi.nlm.nih.gov/pubmed/23762269
http://dx.doi.org/10.1371/journal.pone.0064951
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