title: Expression profiles of glyceraldehyde-3-phosphate dehydrogenase from Clonorchis sinensis: a glycolytic enzyme with plasminogen binding capacity.
abstract: Globally, 15-20 million people are infected with Clonorchis sinensis (C. sinensis) which results in clonorchiasis. In China, clonorchiasis is considered to be one of the fastest-growing food-borne parasitic diseases. That more key molecules of C. sinensis are characterized will be helpful to understand biology and pathogenesis of the carcinogenic liver fluke. Glyceraldehyde-3-phosphate dehydrogenases (GAPDHs) from many species have functions other than their catalytic role in glycolysis. In the present study, we analyzed the sequence and structure of GAPDH from C. sinensis (CsGAPDH) by using bioinformatics tools and obtained its recombinant protein by prokaryotic expression system, to learn its expression profiles and molecular property. CsGAPDH could bind to human intrahepatic biliary epithelial cell in vivo and in vitro by the method of immunofluorescence assays. CsGAPDH also disturbed in lumen of biliary tract near to the parasite in the liver of infected rat. Western blotting analysis together with immunofluorescence assay indicated that CsGAPDH was a component of excretory/secretory proteins (CsESPs) and a surface-localized protein of C. sinensis. Quantitative real-time PCR (Q-PCR) and Western blotting demonstrated that CsGAPDHs are expressed at the life stages of adult worm, metacercaria, and egg, but the expression levels were different from each other. Recombinant CsGAPDH (rCsGAPDH) was confirmed to have the capacity to catalyze the conversion of glyceraldehyde 3-phosphate to D-glycerate 1,3-bisphosphate which was inhibited by AMP in a dose-dependent manner. In addition, rCsGAPDH was able to interact with human plasminogen in a dose-dependent manner by ELISA. The interaction could be inhibited by lysine. The plasminogen binding capacity of rCsGAPDH along with the distribution of CsGAPDH in vivo and in the liver of C. sinensis-infected rat hinted that surface-localized CsGAPDH might play an important role in host invasion of the worm besides its glycolytic activity. Our work will be a cornerstone for getting more messages about CsGAPDH and its role in biology and parasitism of C. sinensis.
authors.affiliation: Department of Clinical Laboratory, The Second People's Hospital of Hefei, Hefei City, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Hu
authors.forename: Erhong
authors.initials: E
authors.lastname: Zhang
authors.forename: Lisi
authors.initials: L
authors.lastname: Huang
authors.forename: Wenfang
authors.initials: W
authors.lastname: Li
authors.forename: Pei
authors.initials: P
authors.lastname: Liang
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002681:China; D003003:Clonorchiasis / Q000201:enzymology / Q000469:parasitology*; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000235:genetics; D019295:Computational Biology; D015971:Gene Expression Regulation, Enzymologic; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015801:Helminth Proteins / Q000302:isolation & purification; D006801:Humans; D008099:Liver / Q000201:enzymology / Q000469:parasitology; D058488:Metacercariae / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D010958:Plasminogen / Q000378:metabolism*; D018510:RNA, Helminth / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D015801:Helminth Proteins; D018510:RNA, Helminth; D011994:Recombinant Proteins; D010958:Plasminogen; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases