title: Cloning and characterization of a selenium-independent glutathione peroxidase (HC29) from adult Haemonchus contortus.
abstract: The complete coding sequence of Haemonchus (H.) contortus HC29 cDNA was generated by rapid amplification of cDNA ends in combination with PCR using primers targeting the 5'- and 3'-ends of the partial mRNA sequence. The cloned HC29 cDNA was shown to be 1,113 bp in size with an open reading frame of 507 bp, encoding a protein of 168 amino acid with a calculated molecular mass of 18.9 kDa. Amino acid sequence analysis revealed that the cloned HC29 cDNA contained the conserved catalytic triad and dimer interface of selenium-independent glutathione peroxidase (GPX). Alignment of the predicted amino acid sequences demonstrated that the protein shared 44.7~80.4% similarity with GPX homologues in the thioredoxin-like family. Phylogenetic analysis revealed close evolutionary proximity of the GPX sequence to the counterpart sequences. These results suggest that HC29 cDNA is a GPX, a member of the thioredoxin-like family. Alignment of the nucleic acid and amino acid sequences of HC29 with those of the reported selenium-independent GPX of H. contortus showed that HC29 contained different types of spliced leader sequences as well as dimer interface sites, although the active sites of both were identical. Enzymatic analysis of recombinant prokaryotic HC29 protein showed activity for the hydrolysis of H(2)O(2). These findings indicate that HC29 is a selenium-independent GPX of H. contortus.
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, Jiangsu, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.forename: Xiaokai
authors.initials: X
authors.lastname: Song
authors.forename: Ruofeng
authors.initials: R
authors.lastname: Yan
authors.forename: Lixin
authors.initials: L
authors.lastname: Xu
authors.forename: Xiangrui
authors.initials: X
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005979:Glutathione Peroxidase / Q000235:genetics* / Q000378:metabolism*; D015511:Goat Diseases / Q000469:parasitology; D006041:Goats; D006188:Haemonchiasis / Q000469:parasitology / Q000517:prevention & control / Q000662:veterinary*; D006189:Haemonchus / Q000201:enzymology* / Q000235:genetics*; D006861:Hydrogen Peroxide / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D018510:RNA, Helminth / Q000737:chemistry / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D051381:Rats; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D018510:RNA, Helminth; D006861:Hydrogen Peroxide; D005979:Glutathione Peroxidase; C080439:selenium-independent glutathione peroxidase