Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9442842 | Experimental Parasitology | 2005 | 8 Pages |
Abstract
The NAD-dependent cytosolic malate dehydrogenase (cMDH, EC 1.1.1.37) plays a pivotal role in the malate-aspartate shuttle pathway that operates in a metabolic coordination between cytosol and mitochondria, and thus is crucial for the survival and pathogenicity of the parasite. In the high throughput sequencing of the cDNA library constructed from the adult stage of Clonorchis sinensis, a cDNA clone containing 1152Â bp insert was identified to encode a putative peptide of 329 amino acids possessing more than 50% amino acid sequence identities with the cMDHs from other organisms such as fish, plant, and mammal. But low sequence similarities have been found between this cMDH and mitochondrial malate dehydrogenase as well as glyoxysomal malate dehydrogenase from other organisms. Northern blot analysis showed the size of the C. sinensis cMDH mRNA was 1.2Â kb. The cMDH was expressed in Escherichia coli M15 as a His-tag fusion protein and purified by BD TALON metal affinity column. The recombinant cMDH showed high MDH activity of 241Â UÂ mgâ1, without lactate dehydrogenase and NADP(H) selectivity. It provides a model for the structure, function analysis, and drug screening on cMDH.
Keywords
Cytosolic malate dehydrogenasecMDHNADPMDHORFClonorchis sinensisIPTGNADPHTrematodeSDS–PAGEβ-nicotinamide adenine dinucleotideβ-nicotinamide adenine dinucleotide phosphatepolyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfateisopropyl-β-d-thiogalactopyranosideHeterologous expressionHeterologous protein expressionProtein sequenceopen reading framelactate dehydrogenaseLDHmalate dehydrogenaseNADNADHpolymerase chain reactionPCR
Related Topics
Life Sciences
Immunology and Microbiology
Parasitology
Authors
Nancai Zheng, Jin Xu, Zhongdao Wu, Jinzhong Chen, Xuchu Hu, Linxia Song, Guang Yang, Chaoneng Ji, Shouyi Chen, Shaohua Gu, Kang Ying, Xinbing Yu,