(19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)

We report a protein-observe (19)F NMR-based ligand titration binding study of human PDI b′x with Δ-somatostatin that also emphasises the need to optimise recombinant protein fluorination when using 5- or 6-fluoroindole. This study highlights a recombinant preference for 5-fluoroindole over 6-fluoroi...

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Autores principales: Curtis-Marof, Rose, Doko, Denisa, Rowe, Michelle L., Richards, Kirsty L., Williamson, Richard A., Howard, Mark J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2014
Materias:
Chemistry
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4157668/
https://www.ncbi.nlm.nih.gov/pubmed/24796794
http://dx.doi.org/10.1039/c4ob00699b
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author Curtis-Marof, Rose
Doko, Denisa
Rowe, Michelle L.
Richards, Kirsty L.
Williamson, Richard A.
Howard, Mark J.
author_facet Curtis-Marof, Rose
Doko, Denisa
Rowe, Michelle L.
Richards, Kirsty L.
Williamson, Richard A.
Howard, Mark J.
author_sort Curtis-Marof, Rose
collection PubMed
description We report a protein-observe (19)F NMR-based ligand titration binding study of human PDI b′x with Δ-somatostatin that also emphasises the need to optimise recombinant protein fluorination when using 5- or 6-fluoroindole. This study highlights a recombinant preference for 5-fluoroindole over 6-fluoroindole; most likely due to the influence of fluorine atomic packing within the folded protein structure. Fluorination affords a single (19)F resonance probe to follow displacement of the protein x-linker as ligand is titrated and provides a dissociation constant of 23 ± 4 μM.
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spelling pubmed-41576682014-09-10 (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI) Curtis-Marof, Rose Doko, Denisa Rowe, Michelle L. Richards, Kirsty L. Williamson, Richard A. Howard, Mark J. Org Biomol Chem Chemistry We report a protein-observe (19)F NMR-based ligand titration binding study of human PDI b′x with Δ-somatostatin that also emphasises the need to optimise recombinant protein fluorination when using 5- or 6-fluoroindole. This study highlights a recombinant preference for 5-fluoroindole over 6-fluoroindole; most likely due to the influence of fluorine atomic packing within the folded protein structure. Fluorination affords a single (19)F resonance probe to follow displacement of the protein x-linker as ligand is titrated and provides a dissociation constant of 23 ± 4 μM. Royal Society of Chemistry 2014-06-21 2014-05-06 /pmc/articles/PMC4157668/ /pubmed/24796794 http://dx.doi.org/10.1039/c4ob00699b Text en This journal is © The Royal Society of Chemistry 2014 http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution 3.0 Unported License (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Chemistry
Curtis-Marof, Rose
Doko, Denisa
Rowe, Michelle L.
Richards, Kirsty L.
Williamson, Richard A.
Howard, Mark J.
(19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)
title (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)
title_full (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)
title_fullStr (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)
title_full_unstemmed (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)
title_short (19)F NMR spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hPDI)
title_sort (19)f nmr spectroscopy monitors ligand binding to recombinantly fluorine-labelled b′x from human protein disulphide isomerase (hpdi)
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4157668/
https://www.ncbi.nlm.nih.gov/pubmed/24796794
http://dx.doi.org/10.1039/c4ob00699b
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