Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen

Ketol-acid reductoisomerase (KARI) orchestrates the biosynthesis of branched-chain amino acids, an elementary reaction in prototrophic organisms as well as a valuable process in biotechnology. Bacterial KARIs belonging to class I organise as dimers or dodecamers and were intensively studied to under...

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Autores principales:	Olivier Nicolas Lemaire, Marie-Caroline Müller, Jörg Kahnt, Tristan Wagner
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	ketol-acid reductoisomerases methanogenic archaea X-ray crystallography conformational rearrangement native purification oligomerisation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/abd59ecacfb74307a3e1bbb35f723d92
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spelling	oai:doaj.org-article:abd59ecacfb74307a3e1bbb35f723d922021-11-25T16:53:44ZStructural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen10.3390/biom111116792218-273Xhttps://doaj.org/article/abd59ecacfb74307a3e1bbb35f723d922021-11-01T00:00:00Zhttps://www.mdpi.com/2218-273X/11/11/1679https://doaj.org/toc/2218-273XKetol-acid reductoisomerase (KARI) orchestrates the biosynthesis of branched-chain amino acids, an elementary reaction in prototrophic organisms as well as a valuable process in biotechnology. Bacterial KARIs belonging to class I organise as dimers or dodecamers and were intensively studied to understand their remarkable specificity towards NADH or NADPH, but also to develop antibiotics. Here, we present the first structural study on a KARI natively isolated from a methanogenic archaea. The dodecameric structure of 0.44-MDa was obtained in two different conformations, an open and close state refined to a resolution of 2.2-Å and 2.1-Å, respectively. These structures illustrate the conformational movement required for substrate and coenzyme binding. While the close state presents the complete NADP bound in front of a partially occupied Mg<sup>2+</sup>-site, the Mg<sup>2+</sup>-free open state contains a tartrate at the nicotinamide location and a bound NADP with the adenine-nicotinamide protruding out of the active site. Structural comparisons show a very high conservation of the active site environment and detailed analyses point towards few specific residues required for the dodecamerisation. These residues are not conserved in other dodecameric KARIs that stabilise their trimeric interface differently, suggesting that dodecamerisation, the cellular role of which is still unknown, might have occurred several times in the evolution of KARIs.Olivier Nicolas LemaireMarie-Caroline MüllerJörg KahntTristan WagnerMDPI AGarticleketol-acid reductoisomerasesmethanogenic archaeaX-ray crystallographyconformational rearrangementnative purificationoligomerisationMicrobiologyQR1-502ENBiomolecules, Vol 11, Iss 1679, p 1679 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	ketol-acid reductoisomerases methanogenic archaea X-ray crystallography conformational rearrangement native purification oligomerisation Microbiology QR1-502
spellingShingle	ketol-acid reductoisomerases methanogenic archaea X-ray crystallography conformational rearrangement native purification oligomerisation Microbiology QR1-502 Olivier Nicolas Lemaire Marie-Caroline Müller Jörg Kahnt Tristan Wagner Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen
description	Ketol-acid reductoisomerase (KARI) orchestrates the biosynthesis of branched-chain amino acids, an elementary reaction in prototrophic organisms as well as a valuable process in biotechnology. Bacterial KARIs belonging to class I organise as dimers or dodecamers and were intensively studied to understand their remarkable specificity towards NADH or NADPH, but also to develop antibiotics. Here, we present the first structural study on a KARI natively isolated from a methanogenic archaea. The dodecameric structure of 0.44-MDa was obtained in two different conformations, an open and close state refined to a resolution of 2.2-Å and 2.1-Å, respectively. These structures illustrate the conformational movement required for substrate and coenzyme binding. While the close state presents the complete NADP bound in front of a partially occupied Mg<sup>2+</sup>-site, the Mg<sup>2+</sup>-free open state contains a tartrate at the nicotinamide location and a bound NADP with the adenine-nicotinamide protruding out of the active site. Structural comparisons show a very high conservation of the active site environment and detailed analyses point towards few specific residues required for the dodecamerisation. These residues are not conserved in other dodecameric KARIs that stabilise their trimeric interface differently, suggesting that dodecamerisation, the cellular role of which is still unknown, might have occurred several times in the evolution of KARIs.
format	article
author	Olivier Nicolas Lemaire Marie-Caroline Müller Jörg Kahnt Tristan Wagner
author_facet	Olivier Nicolas Lemaire Marie-Caroline Müller Jörg Kahnt Tristan Wagner
author_sort	Olivier Nicolas Lemaire
title	Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen
title_short	Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen
title_full	Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen
title_fullStr	Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen
title_full_unstemmed	Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen
title_sort	structural rearrangements of a dodecameric ketol-acid reductoisomerase isolated from a marine thermophilic methanogen
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/abd59ecacfb74307a3e1bbb35f723d92
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Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen

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