A Fe2+-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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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2022.876352/full |
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| author | Luigi Scietti Elisabetta Moroni Daiana Mattoteia Marco Fumagalli Matteo De Marco Lisa Negro Antonella Chiapparino Stefano A. Serapian Francesca De Giorgi Silvia Faravelli Giorgio Colombo Federico Forneris |
| author_facet | Luigi Scietti Elisabetta Moroni Daiana Mattoteia Marco Fumagalli Matteo De Marco Lisa Negro Antonella Chiapparino Stefano A. Serapian Francesca De Giorgi Silvia Faravelli Giorgio Colombo Federico Forneris |
| author_sort | Luigi Scietti |
| collection | DOAJ |
| 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 Fe2+ as simultaneous cofactor and inhibitor of lysyl hydroxylase activity and studied the effect of a promiscuous Fe2+ 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 Fe2+. Together, our results show a fine balance between Fe2+-dependent enzymatic activity and Fe2+-induced self-inhibited states, highlighting exquisite differences between LH/PLODs and related Fe2+, 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. |
| first_indexed | 2024-04-14T02:55:01Z |
| format | Article |
| id | doaj.art-b6a66bc99baa419fbaa8c133faca3ec4 |
| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-04-14T02:55:01Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-b6a66bc99baa419fbaa8c133faca3ec42022-12-22T02:16:08ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2022-08-01910.3389/fmolb.2022.876352876352A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymesLuigi Scietti0Elisabetta Moroni1Daiana Mattoteia2Marco Fumagalli3Matteo De Marco4Lisa Negro5Antonella Chiapparino6Stefano A. Serapian7Francesca De Giorgi8Silvia Faravelli9Giorgio Colombo10Federico Forneris11The Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyConsiglio Nazionale delle Ricerche, Istituto di Scienze e Tecnologie Chimiche “Giulio Natta” (SCITEC-CNR), Milano, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyDepartment of Chemistry, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyDepartment of Chemistry, University of Pavia, Pavia, ItalyThe Armenise-Harvard Laboratory of Structural Biology, Department of Biology and Biotechnology, University of Pavia, Pavia, ItalyMultifunctional 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 Fe2+ as simultaneous cofactor and inhibitor of lysyl hydroxylase activity and studied the effect of a promiscuous Fe2+ 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 Fe2+. Together, our results show a fine balance between Fe2+-dependent enzymatic activity and Fe2+-induced self-inhibited states, highlighting exquisite differences between LH/PLODs and related Fe2+, 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.https://www.frontiersin.org/articles/10.3389/fmolb.2022.876352/fullcollagenlysyl hydroxylase (LH)Fe2+/2-oxoglutarate-dependent dioxygenasesstructure-based drug designmolecular dynamics simulationscancer metastasis |
| spellingShingle | Luigi Scietti Elisabetta Moroni Daiana Mattoteia Marco Fumagalli Matteo De Marco Lisa Negro Antonella Chiapparino Stefano A. Serapian Francesca De Giorgi Silvia Faravelli Giorgio Colombo Federico Forneris A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes Frontiers in Molecular Biosciences collagen lysyl hydroxylase (LH) Fe2+/2-oxoglutarate-dependent dioxygenases structure-based drug design molecular dynamics simulations cancer metastasis |
| title | A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes |
| title_full | A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes |
| title_fullStr | A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes |
| title_full_unstemmed | A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes |
| title_short | A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes |
| title_sort | fe2 dependent self inhibited state influences the druggability of human collagen lysyl hydroxylase lh plod enzymes |
| topic | collagen lysyl hydroxylase (LH) Fe2+/2-oxoglutarate-dependent dioxygenases structure-based drug design molecular dynamics simulations cancer metastasis |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2022.876352/full |
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