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A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms

The dimeric ectonucleotidase CD73 catalyzes the hydrolysis of AMP at the cell surface to form adenosine, a potent suppressor of the immune response. Blocking CD73 activity in the tumor microenvironment can have a beneficial effect on tumor eradication and is a promising approach for cancer therapy....

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Autores principales: Stefano, James E., Lord, Dana M., Zhou, Yanfeng, Jaworski, Julie, Hopke, Joern, Travaline, Tara, Zhang, Ningning, Wong, Karen, Lennon, Amanda, He, Timothy, Bric-Furlong, Eva, Cherrie, Cornishia, Magnay, Tristan, Remy, Elisabeth, Brondyk, William, Qiu, Huawei, Radošević, Katarina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7939394/
https://www.ncbi.nlm.nih.gov/pubmed/33122192
http://dx.doi.org/10.1074/jbc.RA120.012395
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author Stefano, James E.
Lord, Dana M.
Zhou, Yanfeng
Jaworski, Julie
Hopke, Joern
Travaline, Tara
Zhang, Ningning
Wong, Karen
Lennon, Amanda
He, Timothy
Bric-Furlong, Eva
Cherrie, Cornishia
Magnay, Tristan
Remy, Elisabeth
Brondyk, William
Qiu, Huawei
Radošević, Katarina
author_facet Stefano, James E.
Lord, Dana M.
Zhou, Yanfeng
Jaworski, Julie
Hopke, Joern
Travaline, Tara
Zhang, Ningning
Wong, Karen
Lennon, Amanda
He, Timothy
Bric-Furlong, Eva
Cherrie, Cornishia
Magnay, Tristan
Remy, Elisabeth
Brondyk, William
Qiu, Huawei
Radošević, Katarina
author_sort Stefano, James E.
collection PubMed
description The dimeric ectonucleotidase CD73 catalyzes the hydrolysis of AMP at the cell surface to form adenosine, a potent suppressor of the immune response. Blocking CD73 activity in the tumor microenvironment can have a beneficial effect on tumor eradication and is a promising approach for cancer therapy. Biparatopic antibodies binding different regions of CD73 may be a means to antagonize its enzymatic activity. A panel of biparatopic antibodies representing the pairwise combination of 11 parental monoclonal antibodies against CD73 was generated by Fab-arm exchange. Nine variants vastly exceeded the potency of their parental antibodies with ≥90% inhibition of activity and subnanomolar EC(50) values. Pairing the Fabs of parents with nonoverlapping epitopes was both sufficient and necessary whereas monovalent antibodies were poor inhibitors. Some parental antibodies yielded potent biparatopics with multiple partners, one of which (TB19) producing the most potent. The structure of the TB19 Fab with CD73 reveals that it blocks alignment of the N- and C-terminal CD73 domains necessary for catalysis. A separate structure of CD73 with a Fab (TB38) which complements TB19 in a particularly potent biparatopic shows its binding to a nonoverlapping site on the CD73 N-terminal domain. Structural modeling demonstrates a TB19/TB38 biparatopic antibody would be unable to bind the CD73 dimer in a bivalent manner, implicating crosslinking of separate CD73 dimers in its mechanism of action. This ability of a biparatopic antibody to both crosslink CD73 dimers and fix them in an inactive conformation thus represents a highly effective mechanism for the inhibition of CD73 activity.
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spelling pubmed-79393942021-06-08 A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms Stefano, James E. Lord, Dana M. Zhou, Yanfeng Jaworski, Julie Hopke, Joern Travaline, Tara Zhang, Ningning Wong, Karen Lennon, Amanda He, Timothy Bric-Furlong, Eva Cherrie, Cornishia Magnay, Tristan Remy, Elisabeth Brondyk, William Qiu, Huawei Radošević, Katarina J Biol Chem Protein Structure and Folding The dimeric ectonucleotidase CD73 catalyzes the hydrolysis of AMP at the cell surface to form adenosine, a potent suppressor of the immune response. Blocking CD73 activity in the tumor microenvironment can have a beneficial effect on tumor eradication and is a promising approach for cancer therapy. Biparatopic antibodies binding different regions of CD73 may be a means to antagonize its enzymatic activity. A panel of biparatopic antibodies representing the pairwise combination of 11 parental monoclonal antibodies against CD73 was generated by Fab-arm exchange. Nine variants vastly exceeded the potency of their parental antibodies with ≥90% inhibition of activity and subnanomolar EC(50) values. Pairing the Fabs of parents with nonoverlapping epitopes was both sufficient and necessary whereas monovalent antibodies were poor inhibitors. Some parental antibodies yielded potent biparatopics with multiple partners, one of which (TB19) producing the most potent. The structure of the TB19 Fab with CD73 reveals that it blocks alignment of the N- and C-terminal CD73 domains necessary for catalysis. A separate structure of CD73 with a Fab (TB38) which complements TB19 in a particularly potent biparatopic shows its binding to a nonoverlapping site on the CD73 N-terminal domain. Structural modeling demonstrates a TB19/TB38 biparatopic antibody would be unable to bind the CD73 dimer in a bivalent manner, implicating crosslinking of separate CD73 dimers in its mechanism of action. This ability of a biparatopic antibody to both crosslink CD73 dimers and fix them in an inactive conformation thus represents a highly effective mechanism for the inhibition of CD73 activity. American Society for Biochemistry and Molecular Biology 2021-01-13 /pmc/articles/PMC7939394/ /pubmed/33122192 http://dx.doi.org/10.1074/jbc.RA120.012395 Text en © 2020 © 2020 Stefano et al. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protein Structure and Folding
Stefano, James E.
Lord, Dana M.
Zhou, Yanfeng
Jaworski, Julie
Hopke, Joern
Travaline, Tara
Zhang, Ningning
Wong, Karen
Lennon, Amanda
He, Timothy
Bric-Furlong, Eva
Cherrie, Cornishia
Magnay, Tristan
Remy, Elisabeth
Brondyk, William
Qiu, Huawei
Radošević, Katarina
A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
title A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
title_full A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
title_fullStr A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
title_full_unstemmed A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
title_short A highly potent CD73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
title_sort highly potent cd73 biparatopic antibody blocks organization of the enzyme active site through dual mechanisms
topic Protein Structure and Folding
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7939394/
https://www.ncbi.nlm.nih.gov/pubmed/33122192
http://dx.doi.org/10.1074/jbc.RA120.012395
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