Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery

Abstract Oncostatin M (OSM) is a pleiotropic, interleukin-6 family inflammatory cytokine that plays an important role in inflammatory diseases, including inflammatory bowel disease, rheumatoid arthritis, and cancer progression and metastasis. Recently, elevated OSM levels have been found in the seru...

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Autores principales:	Olga A. Mass, Joseph Tuccinardi, Luke Woodbury, Cody L. Wolf, Bri Grantham, Kelsey Holdaway, Xinzhu Pu, Matthew D. King, Don L. Warner, Cheryl L. Jorcyk, Lisa R. Warner
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9d8f0adf525449bc92eb3e4f5ff6c2f0
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spelling	oai:doaj.org-article:9d8f0adf525449bc92eb3e4f5ff6c2f02021-12-02T16:27:54ZBioactive recombinant human oncostatin M for NMR-based screening in drug discovery10.1038/s41598-021-95424-62045-2322https://doaj.org/article/9d8f0adf525449bc92eb3e4f5ff6c2f02021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95424-6https://doaj.org/toc/2045-2322Abstract Oncostatin M (OSM) is a pleiotropic, interleukin-6 family inflammatory cytokine that plays an important role in inflammatory diseases, including inflammatory bowel disease, rheumatoid arthritis, and cancer progression and metastasis. Recently, elevated OSM levels have been found in the serum of COVID-19 patients in intensive care units. Multiple anti-OSM therapeutics have been investigated, but to date no OSM small molecule inhibitors are clinically available. To pursue a high-throughput screening and structure-based drug discovery strategy to design a small molecule inhibitor of OSM, milligram quantities of highly pure, bioactive OSM are required. Here, we developed a reliable protocol to produce highly pure unlabeled and isotope enriched OSM from E. coli for biochemical and NMR studies. High yields (ca. 10 mg/L culture) were obtained in rich and minimal defined media cultures. Purified OSM was characterized by mass spectrometry and circular dichroism. The bioactivity was confirmed by induction of OSM/OSM receptor signaling through STAT3 phosphorylation in human breast cancer cells. Optimized buffer conditions yielded 1H, 15N HSQC NMR spectra with intense, well-dispersed peaks. Titration of 15N OSM with a small molecule inhibitor showed chemical shift perturbations for several key residues with a binding affinity of 12.2 ± 3.9 μM. These results demonstrate the value of bioactive recombinant human OSM for NMR-based small molecule screening.Olga A. MassJoseph TuccinardiLuke WoodburyCody L. WolfBri GranthamKelsey HoldawayXinzhu PuMatthew D. KingDon L. WarnerCheryl L. JorcykLisa R. WarnerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Olga A. Mass Joseph Tuccinardi Luke Woodbury Cody L. Wolf Bri Grantham Kelsey Holdaway Xinzhu Pu Matthew D. King Don L. Warner Cheryl L. Jorcyk Lisa R. Warner Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery
description	Abstract Oncostatin M (OSM) is a pleiotropic, interleukin-6 family inflammatory cytokine that plays an important role in inflammatory diseases, including inflammatory bowel disease, rheumatoid arthritis, and cancer progression and metastasis. Recently, elevated OSM levels have been found in the serum of COVID-19 patients in intensive care units. Multiple anti-OSM therapeutics have been investigated, but to date no OSM small molecule inhibitors are clinically available. To pursue a high-throughput screening and structure-based drug discovery strategy to design a small molecule inhibitor of OSM, milligram quantities of highly pure, bioactive OSM are required. Here, we developed a reliable protocol to produce highly pure unlabeled and isotope enriched OSM from E. coli for biochemical and NMR studies. High yields (ca. 10 mg/L culture) were obtained in rich and minimal defined media cultures. Purified OSM was characterized by mass spectrometry and circular dichroism. The bioactivity was confirmed by induction of OSM/OSM receptor signaling through STAT3 phosphorylation in human breast cancer cells. Optimized buffer conditions yielded 1H, 15N HSQC NMR spectra with intense, well-dispersed peaks. Titration of 15N OSM with a small molecule inhibitor showed chemical shift perturbations for several key residues with a binding affinity of 12.2 ± 3.9 μM. These results demonstrate the value of bioactive recombinant human OSM for NMR-based small molecule screening.
format	article
author	Olga A. Mass Joseph Tuccinardi Luke Woodbury Cody L. Wolf Bri Grantham Kelsey Holdaway Xinzhu Pu Matthew D. King Don L. Warner Cheryl L. Jorcyk Lisa R. Warner
author_facet	Olga A. Mass Joseph Tuccinardi Luke Woodbury Cody L. Wolf Bri Grantham Kelsey Holdaway Xinzhu Pu Matthew D. King Don L. Warner Cheryl L. Jorcyk Lisa R. Warner
author_sort	Olga A. Mass
title	Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery
title_short	Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery
title_full	Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery
title_fullStr	Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery
title_full_unstemmed	Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery
title_sort	bioactive recombinant human oncostatin m for nmr-based screening in drug discovery
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/9d8f0adf525449bc92eb3e4f5ff6c2f0
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Bioactive recombinant human oncostatin M for NMR-based screening in drug discovery

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