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Unraveling the molecular determinants of a rare human mitochondrial disorder caused by the P144L mutation of FDX2
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Orientador(es)
Resumo(s)
Episodic mitochondrial myopathy with or without optic atrophy and reversible leukoencephalopathy (MEOAL) is a rare, orphan autosomal recessive disorder caused by mutations in ferredoxin-2 (FDX2), which is a [2Fe-2S] cluster-binding protein participating in the formation of iron–sulfur clusters in mitochondria. In this biosynthetic pathway, FDX2 works as electron donor to promote the assembly of both [2Fe-2S] and [4Fe-4S] clusters. A recently identified missense mutation of MEOAL is the homozygous mutation c.431C>T (p.P144L) described in six patients from two unrelated families. This mutation alters a highly conserved proline residue located in a loop of FDX2 that is distant from the [2Fe-2S] cluster. How this Pro to Leu substitution damages iron–sulfur cluster biosynthesis is unknown. In this work, we have first compared the structural, dynamic, cluster binding and redox properties of WT and P144L [2Fe-2S] FDX2 to have clues on how the pathogenic P144L mutation can perturb the FDX2 function. Then, we have investigated the interaction of both WT and P144L [2Fe-2S] FDX2 with its physiological electron donor, ferredoxin reductase FDXR, comparing their electron transfer efficiency and protein–protein recognition patterns. Overall, the data indicate that the pathogenic P144L mutation negatively affects the FDXR-dependent electron transfer pathway from NADPH to FDX2, thereby reducing the capacity of FDX2 in assembling both [2Fe-2S] and [4Fe-4S] clusters. Our study also provided solid molecular evidences on the functional role of the C-terminal tail of FDX2 in the electron transfer between FDX2 and FDXR.
Descrição
Funding Information: We would like to thank Dr. Beno\u00EEt D'Autreaux for providing us the plasmids required to produce human FDXR and a detailed protocol to obtain purified FDXR. This work was supported by a grant from Fondazione Telethon and Fondazione CARIPLO (Cariplo Telethon Alliance GJC2021, grant GJC213517) to P.C. and S.C.B. All authors acknowledge support from COST Action FeSImmChemNet CA21115. S.C.B. acknowledges the support of the European Union-NextGenerationEU-National Recovery and Resilience Plan, Mission 4 Component 2-Investment 1.5-THE-Tuscany Health Ecosystem-ECS00000017-CUP B83C22003920001, and of the Italian Ministry of University and Research (MUR) through Dipartimenti di Eccellenza 2023-2027 (DICUS 2.0) to the Department of Chemistry \u201CUgo Schiff\u201D of the University of Florence. Financial support was also provided to R.O.L. by Project MOSTMICRO-ITQB with refs UIDB/04612/2020 and UIDP/04612/2020 and LS4FUTURE Associated Laboratory (LA/P/0087/2020). Funding Information: We would like to thank Dr. Beno\u00EEt D'Autreaux for providing us the plasmids required to produce human FDXR and a detailed protocol to obtain purified FDXR. This work was supported by a grant from Fondazione Telethon and Fondazione CARIPLO (Cariplo Telethon Alliance GJC2021, grant GJC213517) to P.C. and S.C.B. All authors acknowledge support from COST Action FeSImmChemNet CA21115. S.C.B. acknowledges the support of the European Union\u2010NextGenerationEU\u2010National Recovery and Resilience Plan, Mission 4 Component 2\u2010Investment 1.5\u2010THE\u2010Tuscany Health Ecosystem\u2010ECS00000017\u2010CUP B83C22003920001, and of the Italian Ministry of University and Research (MUR) through Dipartimenti di Eccellenza 2023\u20102027 (DICUS 2.0) to the Department of Chemistry \u201CUgo Schiff\u201D of the University of Florence. Financial support was also provided to R.O.L. by Project MOSTMICRO\u2010ITQB with refs UIDB/04612/2020 and UIDP/04612/2020 and LS4FUTURE Associated Laboratory (LA/P/0087/2020). Publisher Copyright: © 2024 The Author(s). Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.
Palavras-chave
FDX2 iron–sulfur protein ISC machinery MEOAL NMR P144L rare disease Biochemistry Molecular Biology