Enzymatic and physico-chemical characteristics of recombinant cMDH and mMDH of Clonorchis sinensis

Zheng, Nancai; Huang, Bin; Xu, Jin; Huang, Shu-Ching; Chen, Jinzhong; Hu, Xuchu; Kong, Ying; Yu, Xinbing

doi:10.1007/s00436-005-0121-8

Cited by 8 publications

(4 citation statements)

References 40 publications

(43 reference statements)

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“…In recent years, our laboratory has constructed a fulllength cDNA plasmid library from C. sinensis adult and established an expression profile comprising 1,775 unigenes (Xu et al 2004), many of which were first cloned and characterized, and most of them were evaluated of their potential for diagnosis Ma et al 2007;Wu et al 2007), vaccine (Huang et al 2007;Zhou et al 2008), and drug target (Zheng et al 2006). Identifying the molecules involve in the development of the parasite and of its pathogenesis is also our important goal.…”

Section: Introductionmentioning

confidence: 99%

Recognition and characterization of TGF-β receptor interacting protein 1 (TRIP-1) containing WD40 repeats from Clonorchis sinensis by bioinformatics, cloning, and expression in Escherichia coli

Zhou

et al. 2008

Parasitol Res

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A cDNA clone encoding a homologue of transforming growth factor beta (TGF-beta) receptor interacting protein 1 (TRIP-1) was recognized and isolated from full-length cDNA plasmid library of Clonorchis sinensis adult. TRIP-1 is a bifunctional molecule in all eukaryote, which modulates the signaling pathway of TGF-beta as a phosphorylation substrate of TGF-beta type II receptor kinase and controls ribosome assembly and mRNA translation as p36 subunit of the eukaryotic translation initiation factor 3. The structural and immunological characteristics of TRIP-1 from C. sinensis (CsTRIP-1) were analyzed by bioinformatics. The complete coding sequence was expressed in Escherichia coli, and the purified recombinant product was obtained. Western blotting with mixed sera from clonorchiasis patients was positive, whereas the normal was negative, suggesting it is a candidate of diagnostic antigen for clonorchiasis. CsTRIP-1 will aid to explore interaction between host and the parasite as well as the mechanism by which TGF-beta controls the development of C. sinensis and participates in the pathogenesis.

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Section: Introductionmentioning

confidence: 99%

Recognition and characterization of TGF-β receptor interacting protein 1 (TRIP-1) containing WD40 repeats from Clonorchis sinensis by bioinformatics, cloning, and expression in Escherichia coli

Zhou

et al. 2008

Parasitol Res

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“…mMDH is a key enzyme in the tricarboxylic acid cycle and in the malate-aspartate shuttle. Zheng et al [78][79][80] have demonstrated that glycolytic enzymes are essential required for the survival and pathogenesis of C. sinensis [78][79][80].…”

Section: Enzymes Of Energy Metabolismmentioning

confidence: 99%

Functional Genes and Proteins of Clonorchis sinensis

Kim

Hong

2009

Korean J Parasitol

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During the past several decades, researches on parasite genetics have progressed from biochemical and serodiagnostic studies to protein chemistry, molecular biology, and functional gene studies. Nowadays, bioinformatics, genomics, and proteomics approaches are being applied by Korean parasitology researchers. As for Clonorchis sinensis, investigations have been carried out to identify its functional genes using forward and reverse genetic approaches and to characterize the biochemical and biological properties of its gene products. The authors review the proteins of cloned genes, which include antigenic proteins, physiologic and metabolic enzymes, and the gene expression profile of Clonorchis sinensis. CHROMOSOMES AND GEOGRAPHICAL ISOLATESC. sinensis has 2n = 56 chromosomes, where n consists of 8 large and 20 small chromosomes. C. sinensis geographical isolates collected in Korea and northeastern China (Liaoning Province) have all demonstrated the same chromosome number, but different karyotypes. Specifically, Korean isolates have 3 metacentric and 1 meta-/submetacentric pairs, whereas Chinese isolates have 2 and 2 pairs, respectively. The 2 isolates have same number, 16 and 8, of submetacentric and subtelomeric pairs [13].Genetic relationships between geographical isolates have been studied by employing sequence analyses of nuclear ribosomal DNA (18S, internal transcribed spacer 1 and 2; ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1 [CO-1]). C. sinensis Korean isolates collected in Kimhae city were compared with Chinese isolates collected in Shenyang, Liaoning and Nanning, and Guangxi provinces. The geographical isolates were nearly identical in terms of nuclear ribosomal and mitochondrial DNA sequences, and revealed no more than a 1% sequence difference between Korean and Chinese isolates [14][15][16][17]. Isozyme electrophoresis had been used to study trematode systematics, and isozymes of the Korean and Chinese isolates show homozygous monomorphic banding patterns. Furthermore, alpha-glycerophosphate dehydrogenase produced a unique band pattern and was found to differentiate geographical isolates of C. sinensis [14]. However, in C. sinensis, intraspecific genetic variations among geographical isolates in the SinoKorea region appear minimal.As compared with O. viverrini, C. sinensis ITS2 revealed 95% identity and differences at 28 nucleotide points. Furthermore, the mitochondrial CO1 gene of O. viverrrini was found to be 96% identical with that of C. sinensis. Even a different genus in the same family Opisthorchidae, namely, O viverrrini appears to be genetically close to C. sinensis [17].

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“…Fumarase (EC 4.2.1.2) from A. suum has been recombinantly expressed and purified in good yield, but no inhibitors of this enzyme have been reported [196]. Similarly, recombinant mitochondrial malate dehydrogenase from C. sinensis has been produced and partially characterised [197,198]. 4,4'-Bisdimethylaminodiphenylcarbinol (Michler's hydrol; Scheme 5) inhibits the mitochondrial enzyme from this species but not the cytoplasmic isoform [198].…”

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“…Similarly, recombinant mitochondrial malate dehydrogenase from C. sinensis has been produced and partially characterised [197,198]. 4,4'-Bisdimethylaminodiphenylcarbinol (Michler's hydrol; Scheme 5) inhibits the mitochondrial enzyme from this species but not the cytoplasmic isoform [198]. The mechanism of this inhibition has not been determined.…”

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confidence: 99%

Metabolic enzymes of helminth parasites: potential as drug targets

Timson¹

2015

CPPS

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Metabolic pathways which extract energy from carbon compounds are essential for an organism's survival. Therefore, inhibition of enzymes in these pathways represents a potential therapeutic strategy to combat parasitic infections. However, the high degree of similarity between host and parasite enzymes makes this strategy potentially difficult. Nevertheless, several existing drugs to treat infections by parasitic helminths (worms) target metabolic enzymes. These include the trivalent antimonials which target phosphofructokinase and Clorsulon which targets phosphoglycerate mutase and phosphoglycerate kinase. Glycolytic enzymes from a variety of helminths have been characterised biochemically, and some inhibitors identified. To date none of these inhibitors have been developed into therapies. Many of these enzymes are externalised from the parasite and so are also of interest in the development of potential vaccines. Less work has been done on tricarboxylic acid cycle enzymes and oxidative phosphorylation complexes. Again, while some inhibitors have been identified none have been developed into drug-like molecules. Barriers to the development of novel drugs targeting metabolic enzymes include the lack of experimentally determined structures of helminth enzymes, lack of direct proof that the enzymes are vital in the parasites and lack of cell culture systems for many helminth species. Nevertheless, the success of Clorsulon (which discriminates between highly similar host and parasite enzymes) should inspire us to consider making serious efforts to discover novel anthelminthics which target metabolic enzymes.

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Enzymatic and physico-chemical characteristics of recombinant cMDH and mMDH of Clonorchis sinensis

Cited by 8 publications

References 40 publications

Recognition and characterization of TGF-β receptor interacting protein 1 (TRIP-1) containing WD40 repeats from Clonorchis sinensis by bioinformatics, cloning, and expression in Escherichia coli

Recognition and characterization of TGF-β receptor interacting protein 1 (TRIP-1) containing WD40 repeats from Clonorchis sinensis by bioinformatics, cloning, and expression in Escherichia coli

Functional Genes and Proteins of Clonorchis sinensis

Metabolic enzymes of helminth parasites: potential as drug targets

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