caa a22 4500 606173749 CHVBK 20210128100734.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s11244-015-0413-x doi (NATIONALLICENCE)springer-10.1007/s11244-015-0413-x Characterization of the [FeFe]-Hydrogenase Maturation Protein HydF by EPR Techniques: Insights into the Catalytic Mechanism [Elektronische Daten] [Marco Albertini, Laura Galazzo, Lorenzo Maso, Francesca Vallese, Paola Berto, Edith De Rosa, Marilena Di Valentin, Paola Costantini, Donatella Carbonera] The catalytic site of [FeFe]-hydrogenase, the "H-cluster”, composed by a [4Fe-4S] unit connected by a cysteinyl residue to a [2Fe] center coordinated by three CO, two CN− and a bridging dithiolate, is assembled in a complex maturation pathway, at present not fully characterized, involving three conserved proteins, HydG, HydE and HydF. In this contribution we review our studies on HydF, a protein which acts as scaffold and carrier for the [2Fe] unit of the H-cluster. HydF is a complex enzyme which contains one [4Fe-4S] cluster binding site, with three conserved cysteine residues and a non-Cys ligand. We have exploited EPR, HYSCORE and PELDOR spectroscopies to get insight into the structure and chemical role of HydF. On the basis of the results we discuss the possibility that the non-Cys ligated Fe atom of the [4Fe-4S] cluster, is the site where the [2Fe] subcluster precursor is anchored and finally processed to be delivered to the hydrogenase (HydA). Our PELDOR experiments on the isolated GTPase domain of HydF, have also suggested that interactions with HydG and HydE proteins may be regulated by the binding of the nucleotide. Springer Science+Business Media New York, 2015 HydF nationallicence Hydrogenase nationallicence EPR nationallicence HYSCORE nationallicence PELDOR nationallicence [4Fe-4S] cluster nationallicence Albertini Marco Department of Chemical Sciences, University of Padova, Via F. Marzolo 1, 35131, Padua, Italy aut Galazzo Laura Department of Chemical Sciences, University of Padova, Via F. Marzolo 1, 35131, Padua, Italy aut Maso Lorenzo Department of Biology, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut Vallese Francesca Department of Biomedical Sciences, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut Berto Paola Department of Biomedical Sciences, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut De Rosa Edith Department of Biology, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut Di Valentin Marilena Department of Chemical Sciences, University of Padova, Via F. Marzolo 1, 35131, Padua, Italy aut Costantini Paola Department of Biology, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut Carbonera Donatella Department of Biomedical Sciences, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut Topics in Catalysis Springer US; http://www.springer-ny.com 58/12-13(2015-09-01), 708-718 1022-5528 58:12-13<708 2015 58 11244 https://doi.org/10.1007/s11244-015-0413-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11244-015-0413-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Albertini Marco Department of Chemical Sciences, University of Padova, Via F. Marzolo 1, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Galazzo Laura Department of Chemical Sciences, University of Padova, Via F. Marzolo 1, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Maso Lorenzo Department of Biology, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Vallese Francesca Department of Biomedical Sciences, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Berto Paola Department of Biomedical Sciences, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut NATIONALLICENCE 700 1- De Rosa Edith Department of Biology, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Di Valentin Marilena Department of Chemical Sciences, University of Padova, Via F. Marzolo 1, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Costantini Paola Department of Biology, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut NATIONALLICENCE 700 1- Carbonera Donatella Department of Biomedical Sciences, University of Padova, Viale G. Colombo 3, 35131, Padua, Italy aut NATIONALLICENCE 773 0- Topics in Catalysis Springer US; http://www.springer-ny.com 58/12-13(2015-09-01), 708-718 1022-5528 58:12-13<708 2015 58 11244