Abstract
Ferritin is an evolutionarily conserved protein that plays a vital role in maintaining iron homeostasis. In this study, we identified a ferritin M (PoFerM) from Japanese flounder (Paralichthys olivaceus) and analyzed its biological property. PoFerM is composed of 176 amino acid residues and contains the conserved ferroxidase diiron center and the ferrihydrite nucleation center typical of M ferritins. Expression of PoFerM occurred in multiple tissues and was most abundant in blood. Bacterial infection upregulated PoFerM expression in head kidney, spleen, and liver in a time-dependent manner. Recombinant PoFerM (rPoFerM) purified from Escherichia coli exhibited iron-chelating activity and inhibited bacterial growth, whereas rPoFerMM, the mutant protein that bears alanine substitution at two conserved residues of the ferroxidase center and the ferrihydrite nucleation center, failed to do so. Oxidative protection analysis showed that rPoFerM, but not rPoFerMM, was able to alleviate the deleterious effect of H2O2-induced free radicals on plasmid DNA and primary flounder cells. Together these results indicate that PoFerM is an iron chelator with antimicrobial and antioxidative properties, all which depend on the conserved ferroxidase center and the ferrihydrite nucleation site.
Similar content being viewed by others
References
Andersen O (1997) Accumulation of waterborne iron and expression of ferritin and transferrin in early developmental stages of brown trout (Salmo trutta). Fish Physiol Biochem 16:223–231
Andersen O, Dehli A, Standal H, Giskegjerde TA, Karstensen R, Rørvik KA (1995) Two ferritin subunits of Atlantic salmon (Salmo salar): cloning of the liver cDNAs and antibody preparation. Mol Mar Biol Biotechnol 4:164–170
Andersen O, Pantopoulos K, Kao HT, Muckenthaler M, Youson JH, Pieribone V (1998) Regulation of iron metabolism in the sanguivore lamprey Lampetra fluviatilis. Molecular cloning of two ferritin subunits and two iron-regulatory proteins (IRP) reveals evolutionary conservation of the iron-regulatory element (IRE)/IRP regulatory system. Eur J Biochem 254:223–229
Arosio P, Ingrassia R, Cavadini P (2009) Ferritins: a family of molecules for iron storage, antioxidation and more. Biochim Biophys Acta 1790:589–599
Barber MF, Elde NC (2014) Nutritional immunity. Escape from bacterial iron piracy through rapid evolution of transferrin. Science 346:1362–1366
Bou-Abdallah F, Papaefthymiou GC, Scheswohl DM, Stanga SD, Arosio P, Chasteen ND (2002) Mu-1, 2-Peroxobridged di-iron (III) dimer formation inhuman H-chain ferritin. Biochem J 364:57–63
Carriquiriborde P, Handy RD, Davies SJ (2004) Physiological modulation of iron metabolism in rainbow trout (Oncorhynchus mykiss) fed low and high iron diets. J Exp Biol 207:75–86
Chen C, Hu YH, Xiao ZZ, Sun L (2013) SmCCL19, a CC chemokine of turbot Scophthalmus maximus, induces leukocyte trafficking and promotes anti-viral and anti-bacterial defense. Fish Shellfish Immunol 35:1677–1682
Crichton RR, Declercq JP (2010) X-ray structures of ferritins and related proteins. Biochim Biophys Acta 1800:706–718
Desjardins LM, Hicks BD, Hilton JW (1987) Iron catalyzed oxidation of trout diets and its effect on the growth and physiological response of rainbow trout. Fish Physiol Biochem 4:173–182
Dunford HB (2002) Oxidations of iron(II)/(III) by hydrogen peroxide: from aquo to enzyme. Coord Chem Rev 233:311–318
Elvitigala DA, Premachandra HK, Whang I, Oh MJ, Jung SJ, Park CJ, Lee J (2013) A teleostean counterpart of ferritin M subunit from rock bream (Oplegnathus fasciatus): an active constituent in iron chelation and DNA protection against oxidative damage, with a modulated expression upon pathogen stress. Fish Shellfish Immunol 35:1455–1465
Elvitigala DA, Priyathilaka TT, Lim BS, Whang I, Yeo SY, Choi CY, Lee J (2014) Molecular profile and functional characterization of the ferritin H subunit from rock bream (Oplegnathus fasciatus), revealing its putative role in host antioxidant and immune defense. Dev Comp Immunol 47:104–114
Giorgi A, Mignogna G, Bellapadrona G, Gattoni M, Chiaraluce R, Consalvi V, Chiancone E, Stefanini S (2008) The unusual co-assembly of H- and M-chains in the ferritin molecule from the Antarctic teleosts Trematomus bernacchii and Trematomus newnesi. Arch Biochem Biophys 478:69–74
Harikrishnan R, Kim JS, Jang IS, Kim MC, Balasundaram C, Heo MS (2011) Molecular characterization and tissue distribution of ferritin M in kelp grouper, Epinephelus bruneus. J Biosci Bioeng 112:541–544
Hentze MW, Rouault TA, Harford JB, Klausner RD (1989) Oxidation-reduction and the molecular mechanism of a regulatory RNA-protein interaction. Science 244:357–359
Hu YH, Zheng WJ, Sun L (2010) Identification and molecular analysis of a ferritin subunit from red drum (Sciaenops ocellatus). Fish Shellfish Immunol 28:678–686
Kwong RWM, Andrés JA, Niyogi S (2010) Molecular evidence and physiological characterization of iron absorption in isolated enterocytes of rainbow trout (Oncorhynchus mykiss): implications for dietary cadmium and lead absorption. Aquat Toxicol 99:343–350
Kwong RWM, Hamilton CD, Niyogi Som (2013) Effects of elevated dietary iron on the gastrointestinal expression of Nramp genes and iron homeostasis in rainbow trout (Oncorhynchus mykiss). Fish Physiol Biochem 39:363–372
Lee JH, Pooley NJ, Mohd-Adnan A, Martin SA (2014) Cloning and characterisation of multiple ferritin isoforms in the Atlantic Salmon (Salmo salar). PLoS ONE 9:e103729
Levi S, Santambrogio P, Cozzi A, Rovida E, Corsi B, Tamborini E, Spada S, Albertini A, Arosio P (1994) The role of the L-chain in ferritin iron incorporation: studies of homo and heteropolymers. J Mol Biol 238:649–654
Liu H, Takano T, Peatman E, Abernathy J, Wang S, Sha Z, Kucuktas H, Xu DH, Klesius P, Liu Z (2010) Molecular characterization and gene expression of the channel catfish ferritin H subunit after bacterial infection and iron treatment. J Exp Zool A Ecol Genet Physiol 313:359–368
Luo Y, Henle ES, Linn S (1996) Oxidative damage to DNA constituents by iron-mediated fenton reactions. The deoxycytidine family. J Biol Chem 30:21167–21176
Mignogna G, Chiaraluce R, Consalvi V, Cavallo S, Stefanini S, Chiancone E (2002) Ferritin from the spleen of the Antarctic teleost Trematomus bernacchii is an M-type homopolymer. Eur J Biochem 269:1600–1606
Orino K, Watanabe K (2008) Molecular, physiological and clinical aspects of the iron storage protein ferritin. Vet J 178:191–201
Orino K, Lehman L, Tsuji Y, Ayaki H, Torti SV, Torti FM (2001) Ferritin and the response to oxidative stress. Biochem J 357:241–247
Reyes-Becerril M, Angulo-Valadez C, Macias ME, Angulo M, Ascencio-Valle F (2014) Iron bioavailability in larvae yellow snapper (Lutjanus argentiventris): cloning and expression analysis of ferritin-H. Fish Shellfish Immunol 37:248–255
Santambrogio P, Levi S, Arosio P, Palagi L, Vecchio G, Lawson DM, Yewdall SJ, Artymiuk PJ, Harrison PM, Jappelli R (1992) Evidence that a salt bridge in the light chain contributes to the physical stability difference between heavy and light human ferritins. J Biol Chem 267:14077–14083
Scudiero R, Esposito MG, Trinchella F (2013) Middle ferritin genes from the icefish Chionodraco rastrospinosus: comparative analysis and evolution of fish ferritins. C R Biol 336:134–141
Shull GE, Theil EC (1982) Translational control of ferritin synthesis by iron in embryonic reticulocytes of the bullfrog. J Biol Chem 257:14187–14191
Skaar EP (2010) The battle for iron between bacterial pathogens and their vertebrate hosts. PLoS Pathog 6(8):e1000949
Theil EC (2007) Coordinating responses to iron and oxygen stress with DNA and mRNA promoters: the ferritin story. Biometals 20:513–521
Theil EC, Eisenstein RS (2000) Combinatorial mRNA regulation: iron regulatory proteins and iso-iron-responsive elements (Iso-IREs). J Biol Chem 275:40659–40662
Wang W, Zhang M, Sun L (2011) Ferritin M of Cynoglossus semilaevis: an iron-binding protein and a broad-spectrum antimicrobial that depends on the integrity of the ferroxidase center and nucleation center for biological activity. Fish Shellfish Immunol 31:269–274
Watt RK (2011) The many faces of the octahedral ferritin protein. Biometals 24:489–500
Yu LP, Hu YH, Sun BG, Sun L (2013) Immunological study of the outer membrane proteins of Vibrio harveyi: insights that link immunoprotectivity to interference with bacterial infection. Fish Shellfish Immunol 35:858–865
Zhang M, Sun K, Sun L (2008) Regulation of autoinducer 2 production and luxS expression in a pathogenic Edwardsiella tarda strain. Microbiology 154:2060–2069
Zhang X, Wei W, Wu H, Xu H, Chang K, Zhang Y (2010) Gene cloning and characterization of ferritin H and M subunits from large yellow croaker (Pseudosciaena crocea). Fish Shellfish Immunol 28:735–742
Zhao G, Bou-Abdallah F, Arosio P, Levi S, Janus-Chandler C, Chasteen ND (2003) Multiple pathways for mineral core formation in mammalian apoferritin. The role of hydrogen peroxide. Biochemistry 42:3142–3150
Zheng WJ, Sun L (2011) Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus). Fish Shellfish Immunol 30:638–645
Zheng WJ, Hu YH, Sun L (2010) Identification and analysis of a Scophthalmusmaximus ferritin that is regulated at transcription level by oxidative stress and bacterial infection. Comp Biochem Physiol 156:222–228
Acknowledgments
This work was supported by the Grants from the 863 High Technology Project of the Chinese Ministry of Science and Technology (2012BAD17B01), the National Natural Science Foundation of China (31330081) and the Taishan Scholar Program of Shandong Province.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Wang, Jj., Sun, L. Ferritin M of Paralichthys olivaceus possesses antimicrobial and antioxidative properties. Fish Physiol Biochem 41, 951–959 (2015). https://doi.org/10.1007/s10695-015-0060-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-015-0060-y