Characterization and localization of Opisthorchis viverrini fructose-1,6-bisphosphate aldolase.
Opisthorchis viverrini (Ov) infection is a long-time public health problem in Thailand that can lead to bile duct cancer, cholangiocarcinoma (CCA). Characterization of the Ov proteins at a molecular level will increase our knowledge of host-parasite interaction that can be applied to new drug, vaccine, or immunodiagnostic development. In this study, an important enzyme in the Ov glycolytic pathway, fructose-1,6-bisphosphate aldolase (FBPA), that had been obtained from a previous study was characterized and immunolocalized. The full-length sequence of OvFBPA gene is 1089bp and encodes 362 amino acids with a predicted molecular weight and isoelectric point of 39.54kDa and 7.61, respectively. Additionally, three OvFBPA isoforms were identified by sequence analysis. The amino acid sequence of OvFBPA-1 characterized in this study shared 98% identity to FBPA isoform 1 of Clonorchis sinensis that was classified based on highly conserved active residues to class-I FBPA. The recombinant OvFBPA-1 protein was expressed as a soluble form in Escherichia coli at 25°C with N-terminal His-tagged fusion protein and the purified OvFBPA-1 protein was used to generate polyclonal antibody in mice. Antibody against rOvFBPA-1 protein was able to detect the native OvFBPA-1 protein in both Ov infected hamster liver section and Ov excretory-secretory (ES) products by immunohistochemistry and western blotting, respectively.
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Jeerati
J
Prompipak
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Thanaset
T
Senawong
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Khuanta
K
Jokchaiyaphum
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Kornpira
K
Siriwes
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Suporn
S
Nuchadomrong
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Thewarach
T
Laha
Tropical Disease Research Laboratory, Department of Pathology, Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Banchob
B
Sripa
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand. Electronic address: gulsiri@kku.ac.th.
Gulsiri
G
Senawong
D000595:Amino Acid Sequence; D000818:Animals; D006224:Cricetinae; D004926:Escherichia coli / Q000235:genetics; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006790:Host-Parasite Interactions; D008099:Liver / Q000469:parasitology*; D051379:Mice; D008813:Mice, Inbred ICR; D009891:Opisthorchis / Q000235:genetics / Q000502:physiology*; D030781:Organisms, Genetically Modified / Q000235:genetics; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D015801:Helminth Proteins; D011994:Recombinant Proteins; D005634:Fructose-Bisphosphate Aldolase
Excretory-secretory antigens; Fructose-1,6-bisphosphate aldolase; Immunolocalization; Opisthorchis viverrini; Protein expression