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A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes

Multifunctional human collagen lysyl hydroxylase (LH/PLOD) enzymes catalyze post-translational hydroxylation and subsequent glycosylation of collagens, enabling their maturation and supramolecular organization in the extracellular matrix (ECM). Recently, the overexpression of LH/PLODs in the tumor m...

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Autores principales: Scietti, Luigi, Moroni, Elisabetta, Mattoteia, Daiana, Fumagalli, Marco, De Marco, Matteo, Negro, Lisa, Chiapparino, Antonella, Serapian, Stefano A., De Giorgi, Francesca, Faravelli, Silvia, Colombo, Giorgio, Forneris, Federico
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9453210/
https://www.ncbi.nlm.nih.gov/pubmed/36090047
http://dx.doi.org/10.3389/fmolb.2022.876352
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author Scietti, Luigi
Moroni, Elisabetta
Mattoteia, Daiana
Fumagalli, Marco
De Marco, Matteo
Negro, Lisa
Chiapparino, Antonella
Serapian, Stefano A.
De Giorgi, Francesca
Faravelli, Silvia
Colombo, Giorgio
Forneris, Federico
author_facet Scietti, Luigi
Moroni, Elisabetta
Mattoteia, Daiana
Fumagalli, Marco
De Marco, Matteo
Negro, Lisa
Chiapparino, Antonella
Serapian, Stefano A.
De Giorgi, Francesca
Faravelli, Silvia
Colombo, Giorgio
Forneris, Federico
author_sort Scietti, Luigi
collection PubMed
description Multifunctional human collagen lysyl hydroxylase (LH/PLOD) enzymes catalyze post-translational hydroxylation and subsequent glycosylation of collagens, enabling their maturation and supramolecular organization in the extracellular matrix (ECM). Recently, the overexpression of LH/PLODs in the tumor microenvironment results in abnormal accumulation of these collagen post-translational modifications, which has been correlated with increased metastatic progression of a wide variety of solid tumors. These observations make LH/PLODs excellent candidates for prospective treatment of aggressive cancers. The recent years have witnessed significant research efforts to facilitate drug discovery on LH/PLODs, including molecular structure characterizations and development of reliable high-throughput enzymatic assays. Using a combination of biochemistry and in silico studies, we characterized the dual role of Fe(2+) as simultaneous cofactor and inhibitor of lysyl hydroxylase activity and studied the effect of a promiscuous Fe(2+) chelating agent, 2,2’-bipyridil, broadly considered a lysyl hydroxylase inhibitor. We found that at low concentrations, 2,2’-bipyridil unexpectedly enhances the LH enzymatic activity by reducing the inhibitory effect of excess Fe(2+). Together, our results show a fine balance between Fe(2+)-dependent enzymatic activity and Fe(2+)-induced self-inhibited states, highlighting exquisite differences between LH/PLODs and related Fe(2+), 2-oxoglutarate dioxygenases and suggesting that conventional structure-based approaches may not be suited for successful inhibitor development. These insights address outstanding questions regarding druggability of LH/PLOD lysyl hydroxylase catalytic site and provide a solid ground for upcoming drug discovery and screening campaigns.
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spelling pubmed-94532102022-09-09 A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes Scietti, Luigi Moroni, Elisabetta Mattoteia, Daiana Fumagalli, Marco De Marco, Matteo Negro, Lisa Chiapparino, Antonella Serapian, Stefano A. De Giorgi, Francesca Faravelli, Silvia Colombo, Giorgio Forneris, Federico Front Mol Biosci Molecular Biosciences Multifunctional human collagen lysyl hydroxylase (LH/PLOD) enzymes catalyze post-translational hydroxylation and subsequent glycosylation of collagens, enabling their maturation and supramolecular organization in the extracellular matrix (ECM). Recently, the overexpression of LH/PLODs in the tumor microenvironment results in abnormal accumulation of these collagen post-translational modifications, which has been correlated with increased metastatic progression of a wide variety of solid tumors. These observations make LH/PLODs excellent candidates for prospective treatment of aggressive cancers. The recent years have witnessed significant research efforts to facilitate drug discovery on LH/PLODs, including molecular structure characterizations and development of reliable high-throughput enzymatic assays. Using a combination of biochemistry and in silico studies, we characterized the dual role of Fe(2+) as simultaneous cofactor and inhibitor of lysyl hydroxylase activity and studied the effect of a promiscuous Fe(2+) chelating agent, 2,2’-bipyridil, broadly considered a lysyl hydroxylase inhibitor. We found that at low concentrations, 2,2’-bipyridil unexpectedly enhances the LH enzymatic activity by reducing the inhibitory effect of excess Fe(2+). Together, our results show a fine balance between Fe(2+)-dependent enzymatic activity and Fe(2+)-induced self-inhibited states, highlighting exquisite differences between LH/PLODs and related Fe(2+), 2-oxoglutarate dioxygenases and suggesting that conventional structure-based approaches may not be suited for successful inhibitor development. These insights address outstanding questions regarding druggability of LH/PLOD lysyl hydroxylase catalytic site and provide a solid ground for upcoming drug discovery and screening campaigns. Frontiers Media S.A. 2022-08-25 /pmc/articles/PMC9453210/ /pubmed/36090047 http://dx.doi.org/10.3389/fmolb.2022.876352 Text en Copyright © 2022 Scietti, Moroni, Mattoteia, Fumagalli, De Marco, Negro, Chiapparino, Serapian, De Giorgi, Faravelli, Colombo and Forneris. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Scietti, Luigi
Moroni, Elisabetta
Mattoteia, Daiana
Fumagalli, Marco
De Marco, Matteo
Negro, Lisa
Chiapparino, Antonella
Serapian, Stefano A.
De Giorgi, Francesca
Faravelli, Silvia
Colombo, Giorgio
Forneris, Federico
A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes
title A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes
title_full A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes
title_fullStr A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes
title_full_unstemmed A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes
title_short A Fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes
title_sort fe(2+)-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (lh/plod) enzymes
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9453210/
https://www.ncbi.nlm.nih.gov/pubmed/36090047
http://dx.doi.org/10.3389/fmolb.2022.876352
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