{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,9]],"date-time":"2025-11-09T07:48:00Z","timestamp":1762674480188,"version":"build-2065373602"},"reference-count":66,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,5,3]],"date-time":"2021-05-03T00:00:00Z","timestamp":1620000000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010661","name":"Horizon 2020 Framework Programme","doi-asserted-by":"publisher","award":["iNEXT-Discovery, Grant Agreement No. 871037"],"award-info":[{"award-number":["iNEXT-Discovery, Grant Agreement No. 871037"]}],"id":[{"id":"10.13039\/100010661","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Multiple mitochondrial dysfunctions syndrome (MMDS) is a rare neurodegenerative disorder associated with mutations in genes with a vital role in the biogenesis of mitochondrial [4Fe\u20134S] proteins. Mutations in one of these genes encoding for BOLA3 protein lead to MMDS type 2 (MMDS2). Recently, a novel phenotype for MMDS2 with complete clinical recovery was observed in a patient containing a novel variant (c.176G &gt; A, p.Cys59Tyr) in compound heterozygosity. In this work, we aimed to rationalize this unique phenotype observed in MMDS2. To do so, we first investigated the structural impact of the Cys59Tyr mutation on BOLA3 by NMR, and then we analyzed how the mutation affects both the formation of a hetero-complex between BOLA3 and its protein partner GLRX5 and the iron\u2013sulfur cluster-binding properties of the hetero-complex by various spectroscopic techniques and by experimentally driven molecular docking. We show that (1) the mutation structurally perturbed the iron\u2013sulfur cluster-binding region of BOLA3, but without abolishing [2Fe\u20132S]2+ cluster-binding on the hetero-complex; (2) tyrosine 59 did not replace cysteine 59 as iron\u2013sulfur cluster ligand; and (3) the mutation promoted the formation of an aberrant apo C59Y BOLA3\u2013GLRX5 complex. All these aspects allowed us to rationalize the unique phenotype observed in MMDS2 caused by Cys59Tyr mutation.<\/jats:p>","DOI":"10.3390\/ijms22094848","type":"journal-article","created":{"date-parts":[[2021,5,5]],"date-time":"2021-05-05T11:06:01Z","timestamp":1620212761000},"page":"4848","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Molecular Basis of Multiple Mitochondrial Dysfunctions Syndrome 2 Caused by CYS59TYR BOLA3 Mutation"],"prefix":"10.3390","volume":"22","author":[{"given":"Giovanni","family":"Saudino","sequence":"first","affiliation":[{"name":"Magnetic Resonance Center (CERM), University of Florence, 50019 Sesto Fiorentino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4140-5413","authenticated-orcid":false,"given":"Dafne","family":"Suraci","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Center (CERM), University of Florence, 50019 Sesto Fiorentino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Veronica","family":"Nasta","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Center (CERM), University of Florence, 50019 Sesto Fiorentino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2376-3321","authenticated-orcid":false,"given":"Simone","family":"Ciofi-Baffoni","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Center (CERM), University of Florence, 50019 Sesto Fiorentino, Italy"},{"name":"Department of Chemistry \u201cUgo Schiff\u201d, University of Florence, 50019 Sesto Fiorentino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0562-5774","authenticated-orcid":false,"given":"Lucia","family":"Banci","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Center (CERM), University of Florence, 50019 Sesto Fiorentino, Italy"},{"name":"Department of Chemistry \u201cUgo Schiff\u201d, University of Florence, 50019 Sesto Fiorentino, Italy"},{"name":"Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine (CIRMMP), 50019 Sesto Fiorentino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1086\/318196","article-title":"A novel syndrome affecting multiple mitochondrial functions, located by microcell-mediated transfer to chromosome 2p14\u20132p13","volume":"68","author":"Seyda","year":"2001","journal-title":"Am. J. Hum. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1146\/annurev-biochem-013118-111540","article-title":"Mechanisms of Mitochondrial Iron-Sulfur Protein Biogenesis","volume":"89","author":"Lill","year":"2020","journal-title":"Annu. Rev. Biochem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1016\/j.tibs.2020.02.001","article-title":"Outlining the Complex Pathway of Mammalian Fe-S Cluster Biogenesis","volume":"45","author":"Maio","year":"2020","journal-title":"Trends Biochem. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1039\/C7MT00180K","article-title":"Iron-sulfur cluster biosynthesis and trafficking\u2014Impact on human disease conditions","volume":"10","author":"Wachnowsky","year":"2018","journal-title":"Metallomics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1016\/j.ajhg.2011.08.011","article-title":"Mutations in iron-sulfur cluster scaffold genes NFU1 and BOLA3 cause a fatal deficiency of multiple respiratory chain and 2-oxoacid dehydrogenase enzymes","volume":"89","author":"Cameron","year":"2011","journal-title":"Am. J. Hum. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1016\/j.ajhg.2011.10.005","article-title":"A fatal mitochondrial disease is associated with defective NFU1 function in the maturation of a subset of mitochondrial Fe-S proteins","volume":"89","author":"Tort","year":"2011","journal-title":"Am. J. Hum. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1093\/brain\/awt328","article-title":"Variant non ketotic hyperglycinemia is caused by mutations in LIAS, BOLA3 and the novel gene GLRX5","volume":"137","author":"Baker","year":"2014","journal-title":"Brain"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2590","DOI":"10.1093\/hmg\/ddt107","article-title":"Mutation of the iron-sulfur cluster assembly gene IBA57 causes severe myopathy and encephalopathy","volume":"22","author":"Ajit","year":"2013","journal-title":"Hum. Mol. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1038\/jhg.2017.35","article-title":"Homozygous p.(Glu87Lys) variant in ISCA1 is associated with a multiple mitochondrial dysfunctions syndrome","volume":"62","author":"Shukla","year":"2017","journal-title":"J. Hum. Genet."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Alfadhel, M. (2019). Multiple Mitochondrial Dysfunctions Syndrome 4 Due to ISCA2 Gene Defects: A Review. Child. Neurol. Open, 6.","DOI":"10.1177\/2329048X19847377"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1007\/s10545-012-9489-7","article-title":"Homozygous missense mutation in BOLA3 causes multiple mitochondrial dysfunctions syndrome in two siblings","volume":"36","author":"Haack","year":"2013","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.ymgme.2017.08.001","article-title":"Impact of mutations within the [Fe-S] cluster or the lipoic acid biosynthesis pathways on mitochondrial protein expression profiles in fibroblasts from patients","volume":"122","author":"Lebigot","year":"2017","journal-title":"Mol. Genet. Metab."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.msard.2018.01.003","article-title":"Mitochondrial leukoencephalopathies: A border zone between acquired and inherited white matter disorders in children?","volume":"20","author":"Bindu","year":"2018","journal-title":"Mult. Scler. Relat. Disord."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kohda, M., Tokuzawa, Y., Kishita, Y., Nyuzuki, H., Moriyama, Y., Mizuno, Y., Hirata, T., Yatsuka, Y., Yamashita-Sugahara, Y., and Nakachi, Y. (2016). A Comprehensive Genomic Analysis Reveals the Genetic Landscape of Mitochondrial Respiratory Chain Complex Deficiencies. PLoS Genet., 12.","DOI":"10.1371\/journal.pgen.1005679"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2787","DOI":"10.1002\/ajmg.a.40490","article-title":"Imaging phenotype of multiple mitochondrial dysfunction syndrome 2, a rare BOLA3-associated leukodystrophy","volume":"176","author":"Nikam","year":"2018","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/jmp.12400","article-title":"Nonketotic hyperglycinemia in captive-bred Vervet monkeys (Chlorocebus aethiops) with cataracts","volume":"48","author":"Khoza","year":"2019","journal-title":"J. Med. Primatol."},{"key":"ref_17","first-page":"100629","article-title":"Pitfalls of relying on genetic testing only to diagnose inherited metabolic disorders in non-western populations\u20145 cases of pyruvate dehydrogenase deficiency from South Africa","volume":"24","author":"Meldau","year":"2020","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/8904_2018_100","article-title":"Severe Leukoencephalopathy with Clinical Recovery Caused by Recessive BOLA3 Mutations","volume":"43","author":"Stutterd","year":"2019","journal-title":"JIMD Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1016\/j.braindev.2018.02.004","article-title":"An infant case of diffuse cerebrospinal lesions and cardiomyopathy caused by a BOLA3 mutation","volume":"40","author":"Nishioka","year":"2018","journal-title":"Brain Dev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.ijcard.2019.01.017","article-title":"Cardiomyopathy in children with mitochondrial disease: Prognosis and genetic background","volume":"279","author":"Kishita","year":"2019","journal-title":"Int. J. Cardiol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Meldau, S., Owen, E.P., Khan, K., and Riordan, G.T. (2020). Mitochondrial molecular genetic results in a South African cohort: Divergent mitochondrial and nuclear DNA findings. J. Clin. Pathol.","DOI":"10.1136\/jclinpath-2020-207026"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2119","DOI":"10.1016\/j.bbagen.2017.05.005","article-title":"Structural insights into the molecular function of human (2Fe-2S) BOLA1-GRX5 and (2Fe-2S) BOLA3-GRX5 complexes","volume":"1861","author":"Nasta","year":"2017","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e16673","DOI":"10.7554\/eLife.16673","article-title":"Mitochondrial Bol1 and Bol3 function as assembly factors for specific iron-sulfur proteins","volume":"5","author":"Uzarska","year":"2016","journal-title":"Elife"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1282","DOI":"10.1039\/C8MT00128F","article-title":"Cluster exchange reactivity of [2Fe-2S] cluster-bridged complexes of BOLA3 with monothiol glutaredoxins","volume":"10","author":"Sen","year":"2018","journal-title":"Metallomics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6724","DOI":"10.1002\/anie.201915615","article-title":"Characterization of [2Fe-2S]-Cluster-Bridged Protein Complexes and Reaction Intermediates by use of Robust Native Mass Spectrometric Methods","volume":"59","author":"Jia","year":"2020","journal-title":"Angew. Chem. Int. Ed. (Engl.)"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2312","DOI":"10.1111\/febs.15140","article-title":"A pathway for assembling [4Fe-4S]2+ clusters in mitochondrial iron-sulfur protein biogenesis","volume":"287","author":"Nasta","year":"2020","journal-title":"FEBS J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"9762","DOI":"10.1073\/pnas.1732541100","article-title":"Subcellular compartmentalization of human Nfu, an iron-sulfur cluster scaffold protein, and its ability to assemble a [4Fe-4S] cluster","volume":"100","author":"Tong","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e15991","DOI":"10.7554\/eLife.15991","article-title":"Role of Nfu1 and Bol3 in iron-sulfur cluster transfer to mitochondrial clients","volume":"5","author":"Melber","year":"2016","journal-title":"Elife"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"166924","DOI":"10.1016\/j.jmb.2021.166924","article-title":"ISCA1 orchestrates ISCA2 and NFU1 in the maturation of human mitochondrial [4Fe-4S] proteins","volume":"433","author":"Suraci","year":"2021","journal-title":"J. Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"123","DOI":"10.3389\/fgene.2015.00123","article-title":"Clinical, biochemical, and genetic spectrum of seven patients with NFU1 deficiency","volume":"6","author":"Ahting","year":"2015","journal-title":"Front. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/s13023-015-0237-6","article-title":"New spastic paraplegia phenotype associated to mutation of NFU1","volume":"10","author":"Tonduti","year":"2015","journal-title":"Orphanet J. Rare Dis"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"412","DOI":"10.3389\/fgene.2014.00412","article-title":"Cavitating leukoencephalopathy with multiple mitochondrial dysfunction syndrome and NFU1 mutations","volume":"5","author":"Invernizzi","year":"2014","journal-title":"Front. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1038\/nprot.2010.32","article-title":"The HADDOCK web server for data-driven biomolecular docking","volume":"5","author":"Bonvin","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1021\/ja026939x","article-title":"HADDOCK: A protein-protein docking approach based on biochemical or biophysical information","volume":"125","author":"Dominguez","year":"2003","journal-title":"J. Am. Chem. Soc."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1810","DOI":"10.1002\/prot.24078","article-title":"Clustering biomolecular complexes by residue contacts similarity","volume":"80","author":"Rodrigues","year":"2012","journal-title":"Proteins"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2316","DOI":"10.1021\/ja00320a017","article-title":"Electronic structure, magnetic properties, ESR and optical spectra for 2-Fe ferredoxin models by LCAO-Xa valence bond theory","volume":"106","author":"Noodleman","year":"1984","journal-title":"J. Am. Chem. Soc."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1021\/bi00168a003","article-title":"Human ferrochelatase is an iron-sulfur protein","volume":"33","author":"Dailey","year":"1994","journal-title":"Biochemistry"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/bs.mie.2017.11.034","article-title":"Biochemical Reconstitution and Spectroscopic Analysis of Iron-Sulfur Proteins","volume":"599","author":"Freibert","year":"2018","journal-title":"Methods Enzym."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"8071","DOI":"10.1021\/bi3006658","article-title":"Spectroscopic and functional characterization of iron-sulfur cluster-bound forms of Azotobacter vinelandii (Nif)IscA","volume":"51","author":"Mapolelo","year":"2012","journal-title":"Biochemistry"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1007\/s00775-018-1573-5","article-title":"The NMR contribution to protein-protein networking in Fe-S protein maturation","volume":"23","author":"Banci","year":"2018","journal-title":"J. Biol. Inorg. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1006\/abbi.1999.1576","article-title":"Electron-nuclear interactions in two prototypical [2Fe-2S] proteins: Selective (chiral) deuteration and analysis of 1H and 2H NMR signals from the alpha and beta hydrogens of cysteinyl residues that ligate the iron in the active sites of human ferredoxin and Anabaena 7120 vegetative ferredoxin","volume":"373","author":"Xia","year":"2000","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1007\/BFb0058197","article-title":"The 1H NMR parameters of magnetically coupled dimers\u2014The Fe2S2 proteins as an example","volume":"72","author":"Banci","year":"1990","journal-title":"Bioinorg. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"4192","DOI":"10.1021\/ic00193a019","article-title":"Effects of phenoxide ligation on iron-sulfur clusters. 2. Preparation and properties of [Fe2S2(OAr)4]2- ions","volume":"23","author":"Cleland","year":"1984","journal-title":"Inorg. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1006\/bbrc.1993.1791","article-title":"Mutated forms of a [2Fe-2S] ferredoxin with serine ligands to the iron-sulfur cluster","volume":"194","author":"Fujinaga","year":"1993","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7188","DOI":"10.1073\/pnas.1302388110","article-title":"X-ray snapshots of possible intermediates in the time course of synthesis and degradation of protein-bound Fe4S4 clusters","volume":"110","author":"Nicolet","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"10124","DOI":"10.1021\/jacs.6b06783","article-title":"Tyrosine-Coordinated P-Cluster in G. diazotrophicus Nitrogenase: Evidence for the Importance of O-Based Ligands in Conformationally Gated Electron Transfer","volume":"138","author":"Owens","year":"2016","journal-title":"J. Am. Chem. Soc."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1042\/bj2430611","article-title":"Tyrosine and tyrosinate fluorescence of pig intestinal Ca2+-binding protein","volume":"243","author":"Hofmann","year":"1987","journal-title":"Biochem. J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/0014-5793(78)80948-X","article-title":"Tyrosinate fluorescence maxima at 345 nm in proteins lacking tryptophan at pH 7","volume":"94","author":"Szabo","year":"1978","journal-title":"FEBS Lett."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1007\/s10895-012-1152-z","article-title":"Fluorescence anisotropy of tyrosinate anion using one-, two- and three-photon excitation: Tyrosinate anion fluorescence","volume":"23","author":"Kierdaszuk","year":"2013","journal-title":"J. Fluoresc."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1021\/bi00302a032","article-title":"Tyrosine and tyrosinate fluorescence of bovine testes calmodulin: Calcium and pH dependence","volume":"23","author":"Pundak","year":"1984","journal-title":"Biochemistry"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1016\/0006-3002(63)91204-6","article-title":"The fluorescence of tyrosine in alkaline solution","volume":"66","author":"Cornog","year":"1963","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1042\/bj0650476","article-title":"Ultraviolet fluorescence of the aromatic amino acids","volume":"65","author":"Teale","year":"1957","journal-title":"Biochem. J."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3809","DOI":"10.1074\/jbc.270.8.3809","article-title":"Fluorescence resolution of the intrinsic tryptophan residues of bovine protein tyrosyl phosphatase","volume":"270","author":"Pokalsky","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"22518","DOI":"10.3390\/ijms151222518","article-title":"Intrinsic tryptophan fluorescence in the detection and analysis of proteins: A focus on F\u00f6rster resonance energy transfer techniques","volume":"15","author":"Ghisaidoobe","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1007\/978-1-4613-9359-7_3","article-title":"Principles of frequency-domain fluorescence spectroscopy and applications to cell membranes","volume":"13","author":"Lakowicz","year":"1988","journal-title":"Sub-Cell. Biochem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"2160","DOI":"10.1021\/ja055971z","article-title":"NMR distinction of single- and multiple-mode binding of small-molecule protein ligands","volume":"128","author":"Reibarkh","year":"2006","journal-title":"J. Am. Chem. Soc."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1016\/j.sbi.2013.07.009","article-title":"Formation of transient protein complexes","volume":"23","author":"Schilder","year":"2013","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"3165","DOI":"10.1093\/hmg\/ddaa172","article-title":"Assembly of the [4Fe-4S] cluster of NFU1 requires the coordinated donation of two [2Fe-2S] clusters from the scaffold proteins, ISCU2 and ISCA1","volume":"29","author":"Jain","year":"2020","journal-title":"Hum. Mol. Genet."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"6203","DOI":"10.1073\/pnas.1400102111","article-title":"[2Fe-2S] cluster transfer in iron-sulfur protein biogenesis","volume":"111","author":"Banci","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"16133","DOI":"10.1021\/jacs.5b10592","article-title":"Elucidating the molecular function of human BOLA2 in GRX3-Dependent anamorsin maturation pathway","volume":"137","author":"Banci","year":"2015","journal-title":"J. Am. Chem. Soc."},{"key":"ref_61","first-page":"79","article-title":"Protein-protein interaction analysis by nuclear magnetic resonance spectroscopy","volume":"261","author":"Gao","year":"2004","journal-title":"Methods Mol. Biol."},{"key":"ref_62","first-page":"128","article-title":"Efficient detection of paramagnetically shifted NMR resonances by optimizing the WEFT pulse sequence","volume":"51","author":"Inubushi","year":"1983","journal-title":"J. Magn. Reson."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1016\/j.jmb.2015.09.014","article-title":"The HADDOCK2.2 Web Server: User-Friendly Integrative Modeling of Biomolecular Complexes","volume":"428","author":"Rodrigues","year":"2016","journal-title":"J. Mol. Biol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1042\/BJ20101286","article-title":"The crystal structure of human GLRX5: Iron-sulfur cluster co-ordination, tetrameric assembly and monomer activity","volume":"433","author":"Johansson","year":"2011","journal-title":"Biochem. J."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1006\/jmbi.1993.1626","article-title":"Comparative protein modelling by satisfaction of spatial restraints","volume":"234","author":"Sali","year":"1993","journal-title":"J. Mol. Biol."},{"key":"ref_66","unstructured":"Case, D.A., Darden, T.A., Cheatham, T.E., Simmerling, C.L., Wang, J., Duke, R.E., Luo, R., Walker, R.C., Zhang, W., and Merz, K.M. (2012). AMBER 12, University of California. [12th ed.]."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4848\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:56:56Z","timestamp":1760162216000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4848"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,3]]},"references-count":66,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["ijms22094848"],"URL":"https:\/\/doi.org\/10.3390\/ijms22094848","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2021,5,3]]}}}