Molecular characterization of an α-N-acetylgalactosaminidase from Clonorchis sinensis

Myoung Ro Lee, Won Gi Yoo, Yu Jung Kim, Dae Won Kim, Shin Hyeong Cho, Kwang Yeon Hwang, Jung Won Ju, Won Ja Lee

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The α-N-acetylgalactosaminidase (α-NAGAL) is an exoglycosidase that selectively cleaves terminal α-linked N-acetylgalactosamines from a variety of sugar chains. A complementary DNA (cDNA) clone encoding a novel Clonorchis sinensis α-NAGAL (Cs-α-NAGAL) was identified in the expressed sequence tags database of the adult C. sinensis liver fluke. The complete coding sequence was 1,308 bp long and encoded a 436-residue protein. The selected glycosidase was manually curated as α-NAGAL (EC 3.2.1.49) based on a composite bioinformatics analysis including a search for orthologues, comparative structure modeling, and the generation of a phylogenetic tree. One orthologue of Cs-α-NAGAL was the Rattus norvegicus α-NAGAL (accession number: NP-001012120) that does not exist in C. sinensis. Cs-α-NAGAL belongs to the GH27 family and the GH-D clan. A phylogenetic analysis revealed that the GH27 family of Cs-α-NAGAL was distinct from GH31 and GH36 within the GH-D clan. The putative 3D structure of Cs-α-NAGAL was built using SWISS-MODEL with a Gallus gallus α-NAGAL template (PDB code 1ktb chain A); this model demonstrated the superimposition of a TIM barrel fold (α/β) structure and substrate binding pocket. Cs-α-NAGAL transcripts were detected in the adult worm and egg cDNA libraries of C. sinensis but not in the metacercaria. Recombinant Cs-α-NAGAL (rCs-α-NAGAL) was expressed in Escherichia coli, and the purified rCs-α-NAGAL was recognized specifically by the C. sinensis-infected human sera. This is the first report of an α-NAGAL protein in the Trematode class, suggesting that it is a potential diagnostic or vaccine candidate with strong antigenicity.

Original languageEnglish
Pages (from-to)2149-2156
Number of pages8
JournalParasitology Research
Volume111
Issue number5
DOIs
Publication statusPublished - 2012 Nov 1

Fingerprint

Clonorchis sinensis
glycosidases
Glycoside Hydrolases
Complementary DNA
Metacercariae
Acetylgalactosamine
liver flukes
Fasciola hepatica
complementary DNA
Gallus gallus
metacercariae
Rattus norvegicus
Expressed Sequence Tags
phylogeny
expressed sequence tags
Computational Biology
Gene Library
Trematoda
bioinformatics
cDNA libraries

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Molecular characterization of an α-N-acetylgalactosaminidase from Clonorchis sinensis. / Lee, Myoung Ro; Yoo, Won Gi; Kim, Yu Jung; Kim, Dae Won; Cho, Shin Hyeong; Hwang, Kwang Yeon; Ju, Jung Won; Lee, Won Ja.

In: Parasitology Research, Vol. 111, No. 5, 01.11.2012, p. 2149-2156.

Research output: Contribution to journalArticle

Lee, MR, Yoo, WG, Kim, YJ, Kim, DW, Cho, SH, Hwang, KY, Ju, JW & Lee, WJ 2012, 'Molecular characterization of an α-N-acetylgalactosaminidase from Clonorchis sinensis', Parasitology Research, vol. 111, no. 5, pp. 2149-2156. https://doi.org/10.1007/s00436-012-3063-y
Lee, Myoung Ro ; Yoo, Won Gi ; Kim, Yu Jung ; Kim, Dae Won ; Cho, Shin Hyeong ; Hwang, Kwang Yeon ; Ju, Jung Won ; Lee, Won Ja. / Molecular characterization of an α-N-acetylgalactosaminidase from Clonorchis sinensis. In: Parasitology Research. 2012 ; Vol. 111, No. 5. pp. 2149-2156.
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