Paragonimus westermani: molecular cloning, expression, and characterization of a recombinant yolk ferritin.
Ferritin is an intracellular protein involved in iron metabolism. A cDNA PwYF-1 cloned from the adult Paragonimus westermani cDNA library encoded a putative polypeptide of 216 amino acids homologous with ferritins of vertebrates and invertebrates. Febinding motifs identified in PwYF-1 polypeptide were conserved and predicted to form a ferroxidase center. PwYF-1 polypeptide contained an extended peptide of 45 amino acids at its C-terminus. Recombinant PwYF-1 protein, expressed and purified from Escherichia coli, showed iron-uptake ability and ferroxidase activity. Ferroxidase activity of recombinant PwYF-1 protein was reactivated by secondary addition of apotransferrin to assay mixture. Mouse immune serum raised against the recombinant PwYF-1 protein recognized specifically 24 kDa protein from adult P. westermani lysate. PwYF-1 protein was localized to vitelline follicles and the eggs of P. westermani. Collectively, PwYF-1 protein was identified as a P. westermani yolk ferritin.
Kim
Tae Yun
TY
Department of Parasitology, Seoul National University College of Medicine, Seoul 110-799, South Korea.
Joo
Il Jung
IJ
Kang
Shin-Yong
SY
Cho
Seung-Yull
SY
Hong
Sung-Jong
SJ
D000595:Amino Acid Sequence; D000818:Animals; D003400:Astacoidea; D001483:Base Sequence; D002570:Ceruloplasmin / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry; D004285:Dogs; D005293:Ferritins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D005786:Gene Expression Regulation; D015151:Immunoblotting; D007150:Immunohistochemistry; D007501:Iron / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010238:Paragonimus / Q000235:genetics* / Q000378:metabolism; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment
D018076:DNA, Complementary; D018509:DNA, Helminth; D011994:Recombinant Proteins; D005293:Ferritins; D007501:Iron; D002570:Ceruloplasmin