Cargando…
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....
Autores principales: | , , , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1783661744344793088 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7939394 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT stefanojamese ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT lorddanam ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT zhouyanfeng ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT jaworskijulie ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT hopkejoern ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT travalinetara ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT zhangningning ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT wongkaren ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT lennonamanda ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT hetimothy ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT bricfurlongeva ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT cherriecornishia ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT magnaytristan ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT remyelisabeth ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT brondykwilliam ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT qiuhuawei ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT radosevickatarina ahighlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT stefanojamese highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT lorddanam highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT zhouyanfeng highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT jaworskijulie highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT hopkejoern highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT travalinetara highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT zhangningning highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT wongkaren highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT lennonamanda highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT hetimothy highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT bricfurlongeva highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT cherriecornishia highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT magnaytristan highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT remyelisabeth highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT brondykwilliam highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT qiuhuawei highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms AT radosevickatarina highlypotentcd73biparatopicantibodyblocksorganizationoftheenzymeactivesitethroughdualmechanisms |