title: Characterization of MIF family proteins: MIF and DDT from rock bream, Oplegnathus fasciatus.
abstract: Macrophage migration inhibitory factor (MIF) is a pleiotropic molecule playing vital roles in various signaling cascades, including cell proliferation, and activation of immune responses against infections. It is well known as a pivotal regulator of innate immunity. In this study, we have rescued and characterized two members of the MIF family, macrophage migration inhibitory factor (OfMIF) and D-Dopachrome tautomerase (OfDDT) from rock bream, Oplegnathus fasciatus. The deduced OfMIF and OfDDT protein sequences revealed the presence of the catalytic oxidoreductase (CXXC), motif. They also possessed highly conserved proline (P(2)) and lysine residues (K(33)), responsible for their isomerase and tautomerase functions. Rock bream MIF and DDT homologues shared higher identity with fish homologues and also with mammals and occupied a distinct position in the phylogenetic tree, depicting their evolutionary conservation. The spatial expression analysis revealed the highest expression of both OfMIF and OfDDT in liver, while portraying constitutive expression in other tissues. The recombinant proteins purified using the Escherichia coli system revealed potent oxidoreductase activity against insulin with both dithiothreitol and glutathione as reducing agents. Stimulation of rock bream head kidney cells with recombinant OfMIF and OfDDT proteins induced the expression of proinflammatory cytokines like tumor necrosis factor alpha (TNF-α), interleukin-8 (IL-8) and interleukin-1β (IL-1β). These results together suggest their involvement in rock bream immune defense and this study on the novel MIF family member DDT from rock bream will pave the way for further studies of this homologue in other teleosts and delineate its multiple functions.
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors.forename: Minyoung
authors.initials: M
authors.lastname: Oh
authors.forename: Saranya Revathy
authors.initials: SR
authors.lastname: Kasthuri
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wan
authors.forename: S D N K
authors.initials: SD
authors.lastname: Bathige
authors.forename: Ilson
authors.initials: I
authors.lastname: Whang
authors.forename: Bong-Soo
authors.initials: BS
authors.lastname: Lim
authors.forename: Hyung-Bok
authors.initials: HB
authors.lastname: Jung
authors.forename: Myung-Joo
authors.initials: MJ
authors.lastname: Oh
authors.forename: Sung-Ju
authors.initials: SJ
authors.lastname: Jung
authors.forename: Sung Yeon
authors.initials: SY
authors.lastname: Kim
authors.forename: Jehee
authors.initials: J
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017756:Aquaculture; D001483:Base Sequence; D003001:Cloning, Molecular; D016207:Cytokines; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D060226:Head Kidney / Q000276:immunology; D019746:Intramolecular Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008263:Macrophage Migration-Inhibitory Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment / Q000662:veterinary
chemical_list: D016207:Cytokines; D018076:DNA, Complementary; D008263:Macrophage Migration-Inhibitory Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase
keywords: D-Dopachrome tautomerase; Expressional analysis; Macrophage migration inhibitory factor; Oxidoreductase assay; Proinflammatory cytokines