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Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors

HSP47 (heat shock protein 47) is a collagen-specific molecular chaperone that is essential for procollagen folding and function. Previous studies have shown that HSP47 binding requires a critical Arg residue at the Y position of the (Gly-Xaa-Yaa) repeats of collagen; however, the exact binding sites...

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Autores principales: Cai, Haiyan, Sasikumar, Parvathy, Little, Gemma, Bihan, Dominique, Hamaia, Samir W., Zhou, Aiwu, Gibbins, Jonathan M., Farndale, Richard W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8301893/
https://www.ncbi.nlm.nih.gov/pubmed/34356607
http://dx.doi.org/10.3390/biom11070983
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author Cai, Haiyan
Sasikumar, Parvathy
Little, Gemma
Bihan, Dominique
Hamaia, Samir W.
Zhou, Aiwu
Gibbins, Jonathan M.
Farndale, Richard W.
author_facet Cai, Haiyan
Sasikumar, Parvathy
Little, Gemma
Bihan, Dominique
Hamaia, Samir W.
Zhou, Aiwu
Gibbins, Jonathan M.
Farndale, Richard W.
author_sort Cai, Haiyan
collection PubMed
description HSP47 (heat shock protein 47) is a collagen-specific molecular chaperone that is essential for procollagen folding and function. Previous studies have shown that HSP47 binding requires a critical Arg residue at the Y position of the (Gly-Xaa-Yaa) repeats of collagen; however, the exact binding sites of HSP47 on native collagens are not fully defined. To address this, we mapped the HSP47 binding sites on collagens through an ELISA binding assay using collagen toolkits, synthetic collagen peptides covering the entire amino acid sequences of collagen types II and III assembled in triple-helical conformation. Our results showed that HSP47 binds to only a few of the GXR motifs in collagen, with most of the HSP47 binding sites identified located near the N-terminal part of the triple-helical region. Molecular modelling and binding energy calculation indicated that residues flanking the key Arg in the collagen sequence also play an important role in defining the high-affinity HSP47 binding site of collagen. Based on this binding mode of HSP47 to collagen, virtual screening targeting both the Arg binding site and its neighboring area on the HSP47 surface, and a subsequent bioassay, we identified two novel compounds with blocking activity towards HSP47 binding of collagen. Overall, our study revealed the native HSP47 binding sites on collagen and provided novel information for the design of small-molecule inhibitors of HSP47.
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spelling pubmed-83018932021-07-24 Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors Cai, Haiyan Sasikumar, Parvathy Little, Gemma Bihan, Dominique Hamaia, Samir W. Zhou, Aiwu Gibbins, Jonathan M. Farndale, Richard W. Biomolecules Article HSP47 (heat shock protein 47) is a collagen-specific molecular chaperone that is essential for procollagen folding and function. Previous studies have shown that HSP47 binding requires a critical Arg residue at the Y position of the (Gly-Xaa-Yaa) repeats of collagen; however, the exact binding sites of HSP47 on native collagens are not fully defined. To address this, we mapped the HSP47 binding sites on collagens through an ELISA binding assay using collagen toolkits, synthetic collagen peptides covering the entire amino acid sequences of collagen types II and III assembled in triple-helical conformation. Our results showed that HSP47 binds to only a few of the GXR motifs in collagen, with most of the HSP47 binding sites identified located near the N-terminal part of the triple-helical region. Molecular modelling and binding energy calculation indicated that residues flanking the key Arg in the collagen sequence also play an important role in defining the high-affinity HSP47 binding site of collagen. Based on this binding mode of HSP47 to collagen, virtual screening targeting both the Arg binding site and its neighboring area on the HSP47 surface, and a subsequent bioassay, we identified two novel compounds with blocking activity towards HSP47 binding of collagen. Overall, our study revealed the native HSP47 binding sites on collagen and provided novel information for the design of small-molecule inhibitors of HSP47. MDPI 2021-07-03 /pmc/articles/PMC8301893/ /pubmed/34356607 http://dx.doi.org/10.3390/biom11070983 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cai, Haiyan
Sasikumar, Parvathy
Little, Gemma
Bihan, Dominique
Hamaia, Samir W.
Zhou, Aiwu
Gibbins, Jonathan M.
Farndale, Richard W.
Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors
title Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors
title_full Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors
title_fullStr Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors
title_full_unstemmed Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors
title_short Identification of HSP47 Binding Site on Native Collagen and Its Implications for the Development of HSP47 Inhibitors
title_sort identification of hsp47 binding site on native collagen and its implications for the development of hsp47 inhibitors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8301893/
https://www.ncbi.nlm.nih.gov/pubmed/34356607
http://dx.doi.org/10.3390/biom11070983
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