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Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference
The human inducible phospho-fructokinase bisphosphatase isoform 3, PFKFB3, is a crucial regulatory node in the cellular metabolism. The enzyme is an important modulator regulating the intracellular fructose-2,6-bisphosphate level. PFKFB3 is a bifunctional enzyme with an exceptionally high kinase to...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6937325/ https://www.ncbi.nlm.nih.gov/pubmed/31889092 http://dx.doi.org/10.1038/s41598-019-56708-0 |
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author | Macut, Helena Hu, Xiao Tarantino, Delia Gilardoni, Ettore Clerici, Francesca Regazzoni, Luca Contini, Alessandro Pellegrino, Sara Luisa Gelmi, Maria |
author_facet | Macut, Helena Hu, Xiao Tarantino, Delia Gilardoni, Ettore Clerici, Francesca Regazzoni, Luca Contini, Alessandro Pellegrino, Sara Luisa Gelmi, Maria |
author_sort | Macut, Helena |
collection | PubMed |
description | The human inducible phospho-fructokinase bisphosphatase isoform 3, PFKFB3, is a crucial regulatory node in the cellular metabolism. The enzyme is an important modulator regulating the intracellular fructose-2,6-bisphosphate level. PFKFB3 is a bifunctional enzyme with an exceptionally high kinase to phosphatase ratio around 740:1. Its kinase activity can be directly inhibited by small molecules acting directly on the kinase active site. On the other hand, here we propose an innovative and indirect strategy for the modulation of PFKFB3 activity, achieved through allosteric bisphosphatase activation. A library of small peptides targeting an allosteric site was discovered and synthesized. The binding affinity was evaluated by microscale thermophoresis (MST). Furthermore, a LC-MS/MS analytical method for assessing the bisphosphatase activity of PFKFB3 was developed. The new method was applied for measuring the activation on bisphosphatase activity with the PFKFB3-binding peptides. The molecular mechanical connection between the newly discovered allosteric site to the bisphosphatase activity was also investigated using both experimental and computational methods. |
format | Online Article Text |
id | pubmed-6937325 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69373252020-01-06 Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference Macut, Helena Hu, Xiao Tarantino, Delia Gilardoni, Ettore Clerici, Francesca Regazzoni, Luca Contini, Alessandro Pellegrino, Sara Luisa Gelmi, Maria Sci Rep Article The human inducible phospho-fructokinase bisphosphatase isoform 3, PFKFB3, is a crucial regulatory node in the cellular metabolism. The enzyme is an important modulator regulating the intracellular fructose-2,6-bisphosphate level. PFKFB3 is a bifunctional enzyme with an exceptionally high kinase to phosphatase ratio around 740:1. Its kinase activity can be directly inhibited by small molecules acting directly on the kinase active site. On the other hand, here we propose an innovative and indirect strategy for the modulation of PFKFB3 activity, achieved through allosteric bisphosphatase activation. A library of small peptides targeting an allosteric site was discovered and synthesized. The binding affinity was evaluated by microscale thermophoresis (MST). Furthermore, a LC-MS/MS analytical method for assessing the bisphosphatase activity of PFKFB3 was developed. The new method was applied for measuring the activation on bisphosphatase activity with the PFKFB3-binding peptides. The molecular mechanical connection between the newly discovered allosteric site to the bisphosphatase activity was also investigated using both experimental and computational methods. Nature Publishing Group UK 2019-12-30 /pmc/articles/PMC6937325/ /pubmed/31889092 http://dx.doi.org/10.1038/s41598-019-56708-0 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Macut, Helena Hu, Xiao Tarantino, Delia Gilardoni, Ettore Clerici, Francesca Regazzoni, Luca Contini, Alessandro Pellegrino, Sara Luisa Gelmi, Maria Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference |
title | Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference |
title_full | Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference |
title_fullStr | Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference |
title_full_unstemmed | Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference |
title_short | Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference |
title_sort | tuning pfkfb3 bisphosphatase activity through allosteric interference |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6937325/ https://www.ncbi.nlm.nih.gov/pubmed/31889092 http://dx.doi.org/10.1038/s41598-019-56708-0 |
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