A macrophage migration inhibitory factor-like tautomerase from Teladorsagia circumcincta (Nematoda: Strongylida).
A macrophage migration inhibitory factor (MIF)-like molecule, Tci-MIF-1, was isolated from Teladorsagia circumcincta and subjected to detailed characterization. A cDNA representing Tci-mif-1 was isolated following its identification in third-stage larvae (L3)-enriched cDNA population. Sequencing of the cDNA indicated a 348-bp open reading frame (ORF) with the closest orthologue being a MIF derived from the human hookworm Ancylostoma ceylanicum. Messenger RNA (mRNA) representing the Tci-MIF-1 transcript was detected in eggs, L3 and adult stages of T. circumcincta. The transcript was also present, but to a lesser extent in fourth-stage larvae (L4). Detection of Tci-MIF-1 protein in T. circumcincta developmental stages reflected the transcript levels identified by reverse transcriptase-PCR. Using immunohistochemistry, the Tci-MIF-1 protein was shown to have a diffuse distribution in L3 tissue, and in L4 and adult stages, the protein was localized to the nematode gut. A recombinant version of Tci-MIF-1 was produced, and enzymic assays indicated that this recombinant protein and a somatic extract of L3 possessed dopachrome tautomerase activity as has been observed previously in other MIF-like molecules. Neither native, purified Tci-MIF nor recombinant Tci-MIF-1 dramatically influenced the in vitro migration of sheep monocytes.
Nisbet
A J
AJ
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Scotland, UK. Alasdair.Nisbet@moredun.ac.uk
Bell
N E V
NE
McNeilly
T N
TN
Knox
D P
DP
Maizels
R M
RM
Meikle
L I
LI
Wildblood
L A
LA
Matthews
J B
JB
D000595:Amino Acid Sequence; D000818:Animals; D002465:Cell Movement; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018509:DNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D041981:Gastrointestinal Tract / Q000737:chemistry; D020869:Gene Expression Profiling; D015801:Helminth Proteins / Q000032:analysis / Q000276:immunology*; D006801:Humans; D007108:Immune Tolerance; D007150:Immunohistochemistry; D019746:Intramolecular Oxidoreductases / Q000032:analysis / Q000276:immunology*; D007814:Larva / Q000737:chemistry; D008264:Macrophages / Q000276:immunology* / Q000469:parasitology; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D014251:Trichostrongyloidea / Q000737:chemistry / Q000201:enzymology* / Q000276:immunology*
D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase