{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Wu Y"],"funding":["National Natural Science Foundation of China","General project of Zhejiang Provincial Education Department","K.C. Wong Magna Fund at Ningbo University"],"pagination":["5378"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9785301"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["14(24)"],"pubmed_abstract":["Ferritin is widely acknowledged as a conservative iron storage protein found in almost all living kingdoms. <i>Apostichopus japonicus</i> (Selenka) is among the oldest echinoderm fauna and has unique regenerative potential, but the catalytic mechanism of iron oxidation in <i>A. japonicus</i> ferritin (AjFER) remains elusive. We previously identified several potential metal-binding sites at the ferroxidase center, the three- and four-fold channels in AjFER. Herein, we prepared AjFER, AjFER-E25A/E60A/E105A, AjFER-D129A/E132A, and AjFER-E168A mutants, investigated their structures, and functionally characterized these ferritins with respect to Fe<sup>2+</sup> uptake using X-ray techniques together with biochemical analytical methods. A crystallographic model of the AjFER-D129A/E132A mutant, which was solved to a resolution of 1.98 Å, suggested that the substitutions had a significant influence on the quaternary structure of the three-fold channel compared to that of AjFER. The structures of these ferritins in solution were determined based on the molecular envelopes of AjFER and its variants by small-angle X-ray scattering, and the structures were almost consistent with the characteristics of well-folded and globular-shaped proteins. Comparative biochemical analyses indicated that site-directed mutagenesis of metal-binding sites in AjFER presented relatively low rates of iron oxidation and thermostability, as well as weak iron-binding affinity, suggesting that these potential metal-binding sites play critical roles in the catalytic activity of ferritin. These findings provide profound insight into the structure-function relationships related to marine invertebrate ferritins."],"journal":["Polymers"],"pubmed_title":["Structural and Functional Insights into the Roles of Potential Metal-Binding Sites in <i>Apostichopus japonicus</i> Ferritin."],"pmcid":["PMC9785301"],"funding_grant_id":["Y202146277","41676159"],"pubmed_authors":["Su C","Feng Y","Huo C","Lu C","Ming T","Liu Y","Li Y","Qiu X","Zhang Z","Han J","Su X","Wu Y","Zhou J"],"additional_accession":[]},"is_claimable":false,"name":"Structural and Functional Insights into the Roles of Potential Metal-Binding Sites in <i>Apostichopus japonicus</i> Ferritin.","description":"Ferritin is widely acknowledged as a conservative iron storage protein found in almost all living kingdoms. <i>Apostichopus japonicus</i> (Selenka) is among the oldest echinoderm fauna and has unique regenerative potential, but the catalytic mechanism of iron oxidation in <i>A. japonicus</i> ferritin (AjFER) remains elusive. We previously identified several potential metal-binding sites at the ferroxidase center, the three- and four-fold channels in AjFER. Herein, we prepared AjFER, AjFER-E25A/E60A/E105A, AjFER-D129A/E132A, and AjFER-E168A mutants, investigated their structures, and functionally characterized these ferritins with respect to Fe<sup>2+</sup> uptake using X-ray techniques together with biochemical analytical methods. A crystallographic model of the AjFER-D129A/E132A mutant, which was solved to a resolution of 1.98 Å, suggested that the substitutions had a significant influence on the quaternary structure of the three-fold channel compared to that of AjFER. The structures of these ferritins in solution were determined based on the molecular envelopes of AjFER and its variants by small-angle X-ray scattering, and the structures were almost consistent with the characteristics of well-folded and globular-shaped proteins. Comparative biochemical analyses indicated that site-directed mutagenesis of metal-binding sites in AjFER presented relatively low rates of iron oxidation and thermostability, as well as weak iron-binding affinity, suggesting that these potential metal-binding sites play critical roles in the catalytic activity of ferritin. These findings provide profound insight into the structure-function relationships related to marine invertebrate ferritins.","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Dec","modification":"2026-04-08T18:24:13.105Z","creation":"2026-04-08T09:14:23.108Z"},"accession":"S-EPMC9785301","cross_references":{"pubmed":["36559745"],"doi":["10.3390/polym14245378"]}}