Identification and biochemical characterization of macrophage migration inhibitory factor-2 (MIF-2) homologue of human lymphatic filarial parasite, Wuchereria bancrofti.
Identifieur interne : 001C96 ( Main/Exploration ); précédent : 001C95; suivant : 001C97Identification and biochemical characterization of macrophage migration inhibitory factor-2 (MIF-2) homologue of human lymphatic filarial parasite, Wuchereria bancrofti.
Auteurs : Nikhil Chauhan [Inde] ; Rohit Sharma [Inde] ; S L Hoti [Inde]Source :
- Acta tropica [ 1873-6254 ] ; 2015.
Descripteurs français
- KwdFr :
- ADN complémentaire (génétique), Alignement de séquences, Animaux, Données de séquences moléculaires, Facteurs inhibiteurs de la migration des macrophages (), Facteurs inhibiteurs de la migration des macrophages (génétique), Facteurs inhibiteurs de la migration des macrophages (métabolisme), Filariose lymphatique (parasitologie), Humains, Phylogénie, Séquence d'acides aminés, Wuchereria bancrofti (génétique), Wuchereria bancrofti (métabolisme).
- MESH :
- génétique : ADN complémentaire, Facteurs inhibiteurs de la migration des macrophages, Wuchereria bancrofti.
- métabolisme : Facteurs inhibiteurs de la migration des macrophages, Wuchereria bancrofti.
- parasitologie : Filariose lymphatique.
- Alignement de séquences, Animaux, Données de séquences moléculaires, Facteurs inhibiteurs de la migration des macrophages, Humains, Phylogénie, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, DNA, Complementary (genetics), Elephantiasis, Filarial (parasitology), Humans, Macrophage Migration-Inhibitory Factors (chemistry), Macrophage Migration-Inhibitory Factors (genetics), Macrophage Migration-Inhibitory Factors (metabolism), Molecular Sequence Data, Phylogeny, Sequence Alignment, Wuchereria bancrofti (genetics), Wuchereria bancrofti (metabolism).
- MESH :
- chemical , chemistry : Macrophage Migration-Inhibitory Factors.
- chemical , genetics : DNA, Complementary, Macrophage Migration-Inhibitory Factors.
- chemical , metabolism : Macrophage Migration-Inhibitory Factors.
- genetics : Wuchereria bancrofti.
- metabolism : Wuchereria bancrofti.
- parasitology : Elephantiasis, Filarial.
- Amino Acid Sequence, Animals, Humans, Molecular Sequence Data, Phylogeny, Sequence Alignment.
Abstract
Homologues of human macrophage migration inhibitory factor (hMIF) have been reported from vertebrates, invertebrates and prokaryotes, as well as plants. Filarial parasites produce two homologues of hMIF viz., MIF-1 and MIF-2, which play important role in the host immune modulation. Earlier, we have characterized MIF-1 (Wba-mif-1) from Wuchereria bancrofti, the major causal organism of human lymphatic filariasis. Here, we are reporting the molecular and biochemical characterization of MIF-2 from this parasite (Wba-mif-2). The complete Wba-mif-2 gene and its cDNA were amplified, cloned and sequenced. The size of Wba-mif-2 gene and cDNA were found to be 4.275 kb and 363 bp, respectively. The gene annotation revealed the presence of a large intron of 3.912 kb interspersed with two exons of 183 bp and 180 bp. The alignment of derived amino acid sequences of Wba-MIF-2 with Wba-MIF-1 showed 44% homology. The conserved CXXC oxido-reductase catalytic site present in Wba-mif-1 was found absent in Wba-mif-2 coding sequence. The amplified Wba-mif-2 cDNA was cloned into an expression vector pRSET-B and transformed into salt inducible Escherichia coli strain GJ1158. The expressed recombinant Wba-MIF-2 protein showed tautomerase activity against L-dopachrome methyl ester and the specific activity was determined to be 18.57±0.77 μmol/mg/min. Three known inhibitors of hMIF tautomerase activity significantly inhibited the tautomerase activity of recombinant Wba-MIF-2. Although the conserved CXXC oxido-reductase motif is absent in Wba-mif-2, the recombinant protein showed significant oxido-reductase activity in the insulin reduction assay, possibly because of the presence of vicinal cysteine residues.
DOI: 10.1016/j.actatropica.2014.10.009
PubMed: 25446175
Affiliations:
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Electronic address: slhoti@yahoo.com.</nlm:affiliation><country xml:lang="fr">Inde</country><wicri:regionArea>Regional Medical Research Centre (ICMR), Nehru Nagar, Belgaum, Karnataka</wicri:regionArea><wicri:noRegion>Karnataka</wicri:noRegion></affiliation></author></analytic><series><title level="j">Acta tropica</title><idno type="eISSN">1873-6254</idno><imprint><date when="2015" type="published">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>DNA, Complementary (genetics)</term><term>Elephantiasis, Filarial (parasitology)</term><term>Humans</term><term>Macrophage Migration-Inhibitory Factors (chemistry)</term><term>Macrophage Migration-Inhibitory Factors (genetics)</term><term>Macrophage Migration-Inhibitory Factors (metabolism)</term><term>Molecular Sequence Data</term><term>Phylogeny</term><term>Sequence Alignment</term><term>Wuchereria bancrofti (genetics)</term><term>Wuchereria bancrofti (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (génétique)</term><term>Alignement de séquences</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Facteurs inhibiteurs de la migration des macrophages ()</term><term>Facteurs inhibiteurs de la migration des macrophages (génétique)</term><term>Facteurs inhibiteurs de la migration des macrophages (métabolisme)</term><term>Filariose lymphatique (parasitologie)</term><term>Humains</term><term>Phylogénie</term><term>Séquence d'acides aminés</term><term>Wuchereria bancrofti (génétique)</term><term>Wuchereria bancrofti (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Macrophage Migration-Inhibitory Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term><term>Macrophage Migration-Inhibitory Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Macrophage Migration-Inhibitory Factors</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term><term>Facteurs inhibiteurs de la migration des macrophages</term><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Facteurs inhibiteurs de la migration des macrophages</term><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="parasitologie" xml:lang="fr"><term>Filariose lymphatique</term></keywords><keywords scheme="MESH" qualifier="parasitology" xml:lang="en"><term>Elephantiasis, Filarial</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Humans</term><term>Molecular Sequence Data</term><term>Phylogeny</term><term>Sequence Alignment</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Alignement de séquences</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Facteurs inhibiteurs de la migration des macrophages</term><term>Humains</term><term>Phylogénie</term><term>Séquence d'acides aminés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Homologues of human macrophage migration inhibitory factor (hMIF) have been reported from vertebrates, invertebrates and prokaryotes, as well as plants. Filarial parasites produce two homologues of hMIF viz., MIF-1 and MIF-2, which play important role in the host immune modulation. Earlier, we have characterized MIF-1 (Wba-mif-1) from Wuchereria bancrofti, the major causal organism of human lymphatic filariasis. Here, we are reporting the molecular and biochemical characterization of MIF-2 from this parasite (Wba-mif-2). The complete Wba-mif-2 gene and its cDNA were amplified, cloned and sequenced. The size of Wba-mif-2 gene and cDNA were found to be 4.275 kb and 363 bp, respectively. The gene annotation revealed the presence of a large intron of 3.912 kb interspersed with two exons of 183 bp and 180 bp. The alignment of derived amino acid sequences of Wba-MIF-2 with Wba-MIF-1 showed 44% homology. The conserved CXXC oxido-reductase catalytic site present in Wba-mif-1 was found absent in Wba-mif-2 coding sequence. The amplified Wba-mif-2 cDNA was cloned into an expression vector pRSET-B and transformed into salt inducible Escherichia coli strain GJ1158. The expressed recombinant Wba-MIF-2 protein showed tautomerase activity against L-dopachrome methyl ester and the specific activity was determined to be 18.57±0.77 μmol/mg/min. Three known inhibitors of hMIF tautomerase activity significantly inhibited the tautomerase activity of recombinant Wba-MIF-2. Although the conserved CXXC oxido-reductase motif is absent in Wba-mif-2, the recombinant protein showed significant oxido-reductase activity in the insulin reduction assay, possibly because of the presence of vicinal cysteine residues.</div></front></TEI><affiliations><list><country><li>Inde</li></country></list><tree><country name="Inde"><noRegion><name sortKey="Chauhan, Nikhil" sort="Chauhan, Nikhil" uniqKey="Chauhan N" first="Nikhil" last="Chauhan">Nikhil Chauhan</name></noRegion><name sortKey="Hoti, S L" sort="Hoti, S L" uniqKey="Hoti S" first="S L" last="Hoti">S L Hoti</name><name sortKey="Sharma, Rohit" sort="Sharma, Rohit" uniqKey="Sharma R" first="Rohit" last="Sharma">Rohit Sharma</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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