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Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates

Human growth hormone (hGH) is a pituitary-derived endocrine protein that regulates several critical postnatal physiologic processes including growth, organ development, and metabolism. Following adulthood, GH is also a regulator of multiple pathologies like fibrosis, cancer, and diabetes. Therefore,...

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Autores principales: Basu, Reetobrata, Brody, Rich, Sandbhor, Uday, Kulkarni, Prateek, Davis, Emily, Swegan, Deborah, Caggiano, Lydia J., Brenya, Edward, Neggers, Sebastian, Kopchick, John J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10410519/
https://www.ncbi.nlm.nih.gov/pubmed/37442239
http://dx.doi.org/10.1016/j.jbc.2023.105030
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author Basu, Reetobrata
Brody, Rich
Sandbhor, Uday
Kulkarni, Prateek
Davis, Emily
Swegan, Deborah
Caggiano, Lydia J.
Brenya, Edward
Neggers, Sebastian
Kopchick, John J.
author_facet Basu, Reetobrata
Brody, Rich
Sandbhor, Uday
Kulkarni, Prateek
Davis, Emily
Swegan, Deborah
Caggiano, Lydia J.
Brenya, Edward
Neggers, Sebastian
Kopchick, John J.
author_sort Basu, Reetobrata
collection PubMed
description Human growth hormone (hGH) is a pituitary-derived endocrine protein that regulates several critical postnatal physiologic processes including growth, organ development, and metabolism. Following adulthood, GH is also a regulator of multiple pathologies like fibrosis, cancer, and diabetes. Therefore, there is a significant pharmaceutical interest in developing antagonists of hGH action. Currently, there is a single FDA-approved antagonist of the hGH receptor (hGHR) prescribed for treating patients with acromegaly and discovered in our laboratory almost 3 decades ago. Here, we present the first data on the structure and function of a new set of protein antagonists with the full range of hGH actions—dual antagonists of hGH binding to the GHR as well as that of hGH binding to the prolactin receptor. We describe the site-specific PEG conjugation, purification, and subsequent characterization using MALDI-TOF, size-exclusion chromatography, thermostability, and biochemical activity in terms of ELISA-based binding affinities with GHR and prolactin receptor. Moreover, these novel hGHR antagonists display distinct antagonism of GH-induced GHR intracellular signaling in vitro and marked reduction in hepatic insulin-like growth factor 1 output in vivo. Lastly, we observed potent anticancer biological efficacies of these novel hGHR antagonists against human cancer cell lines. In conclusion, we propose that these new GHR antagonists have potential for development towards multiple clinical applications related to GH-associated pathologies.
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spelling pubmed-104105192023-08-10 Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates Basu, Reetobrata Brody, Rich Sandbhor, Uday Kulkarni, Prateek Davis, Emily Swegan, Deborah Caggiano, Lydia J. Brenya, Edward Neggers, Sebastian Kopchick, John J. J Biol Chem Research Article Human growth hormone (hGH) is a pituitary-derived endocrine protein that regulates several critical postnatal physiologic processes including growth, organ development, and metabolism. Following adulthood, GH is also a regulator of multiple pathologies like fibrosis, cancer, and diabetes. Therefore, there is a significant pharmaceutical interest in developing antagonists of hGH action. Currently, there is a single FDA-approved antagonist of the hGH receptor (hGHR) prescribed for treating patients with acromegaly and discovered in our laboratory almost 3 decades ago. Here, we present the first data on the structure and function of a new set of protein antagonists with the full range of hGH actions—dual antagonists of hGH binding to the GHR as well as that of hGH binding to the prolactin receptor. We describe the site-specific PEG conjugation, purification, and subsequent characterization using MALDI-TOF, size-exclusion chromatography, thermostability, and biochemical activity in terms of ELISA-based binding affinities with GHR and prolactin receptor. Moreover, these novel hGHR antagonists display distinct antagonism of GH-induced GHR intracellular signaling in vitro and marked reduction in hepatic insulin-like growth factor 1 output in vivo. Lastly, we observed potent anticancer biological efficacies of these novel hGHR antagonists against human cancer cell lines. In conclusion, we propose that these new GHR antagonists have potential for development towards multiple clinical applications related to GH-associated pathologies. American Society for Biochemistry and Molecular Biology 2023-07-11 /pmc/articles/PMC10410519/ /pubmed/37442239 http://dx.doi.org/10.1016/j.jbc.2023.105030 Text en © 2023 The Authors 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 Research Article
Basu, Reetobrata
Brody, Rich
Sandbhor, Uday
Kulkarni, Prateek
Davis, Emily
Swegan, Deborah
Caggiano, Lydia J.
Brenya, Edward
Neggers, Sebastian
Kopchick, John J.
Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates
title Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates
title_full Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates
title_fullStr Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates
title_full_unstemmed Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates
title_short Structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific PEG conjugates
title_sort structure and function of a dual antagonist of the human growth hormone and prolactin receptors with site-specific peg conjugates
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10410519/
https://www.ncbi.nlm.nih.gov/pubmed/37442239
http://dx.doi.org/10.1016/j.jbc.2023.105030
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