Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch

Abstract [FeFe]-hydrogenases catalyse the reduction of protons to hydrogen at a complex 2Fe[4Fe4S] center called H-cluster. The assembly of this active site is a multistep process involving three proteins, HydE, HydF and HydG. According to the current models, HydF has the key double role of scaffold...

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Autores principales:	Laura Galazzo, Lorenzo Maso, Edith De Rosa, Marco Bortolus, Davide Doni, Laura Acquasaliente, Vincenzo De Filippis, Paola Costantini, Donatella Carbonera
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d59cbdba85a14acba716179d46b1d7cf
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spelling	oai:doaj.org-article:d59cbdba85a14acba716179d46b1d7cf2021-12-02T16:06:43ZIdentifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch10.1038/s41598-017-01886-y2045-2322https://doaj.org/article/d59cbdba85a14acba716179d46b1d7cf2017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01886-yhttps://doaj.org/toc/2045-2322Abstract [FeFe]-hydrogenases catalyse the reduction of protons to hydrogen at a complex 2Fe[4Fe4S] center called H-cluster. The assembly of this active site is a multistep process involving three proteins, HydE, HydF and HydG. According to the current models, HydF has the key double role of scaffold, upon which the final H-cluster precursor is assembled, and carrier to transfer it to the target hydrogenase. The X-ray structure of HydF indicates that the protein is a homodimer with both monomers carrying two functional domains: a C-terminal FeS cluster-binding domain, where the precursor is assembled, and a N-terminal GTPase domain, whose exact contribution to cluster biogenesis and hydrogenase activation is still elusive. We previously obtained several hints suggesting that the binding of GTP to HydF could be involved in the interactions of this scaffold protein with the other maturases and with the hydrogenase itself. In this work, by means of site directed spin labeling coupled to EPR/PELDOR spectroscopy, we explored the conformational changes induced in a recombinant HydF protein by GTP binding, and provide the first clue that the HydF GTPase domain could be involved in the H-cluster assembly working as a molecular switch similarly to other known small GTPases.Laura GalazzoLorenzo MasoEdith De RosaMarco BortolusDavide DoniLaura AcquasalienteVincenzo De FilippisPaola CostantiniDonatella CarboneraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-14 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Laura Galazzo Lorenzo Maso Edith De Rosa Marco Bortolus Davide Doni Laura Acquasaliente Vincenzo De Filippis Paola Costantini Donatella Carbonera Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch
description	Abstract [FeFe]-hydrogenases catalyse the reduction of protons to hydrogen at a complex 2Fe[4Fe4S] center called H-cluster. The assembly of this active site is a multistep process involving three proteins, HydE, HydF and HydG. According to the current models, HydF has the key double role of scaffold, upon which the final H-cluster precursor is assembled, and carrier to transfer it to the target hydrogenase. The X-ray structure of HydF indicates that the protein is a homodimer with both monomers carrying two functional domains: a C-terminal FeS cluster-binding domain, where the precursor is assembled, and a N-terminal GTPase domain, whose exact contribution to cluster biogenesis and hydrogenase activation is still elusive. We previously obtained several hints suggesting that the binding of GTP to HydF could be involved in the interactions of this scaffold protein with the other maturases and with the hydrogenase itself. In this work, by means of site directed spin labeling coupled to EPR/PELDOR spectroscopy, we explored the conformational changes induced in a recombinant HydF protein by GTP binding, and provide the first clue that the HydF GTPase domain could be involved in the H-cluster assembly working as a molecular switch similarly to other known small GTPases.
format	article
author	Laura Galazzo Lorenzo Maso Edith De Rosa Marco Bortolus Davide Doni Laura Acquasaliente Vincenzo De Filippis Paola Costantini Donatella Carbonera
author_facet	Laura Galazzo Lorenzo Maso Edith De Rosa Marco Bortolus Davide Doni Laura Acquasaliente Vincenzo De Filippis Paola Costantini Donatella Carbonera
author_sort	Laura Galazzo
title	Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch
title_short	Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch
title_full	Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch
title_fullStr	Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch
title_full_unstemmed	Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch
title_sort	identifying conformational changes with site-directed spin labeling reveals that the gtpase domain of hydf is a molecular switch
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/d59cbdba85a14acba716179d46b1d7cf
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Identifying conformational changes with site-directed spin labeling reveals that the GTPase domain of HydF is a molecular switch

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