title: Glyceraldehyde-3-phosphate dehydrogenase from Chironomidae showed differential activity towards metals.
abstract: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is known to interact with different biomolecules and was implicated in many novel cellular activities including programmed cell death, nuclear RNA transport unrelated to the commonly known carbohydrate metabolism. We reported here the purification of GAPDH from Chironomidae larvae (Insecta, Diptera) that showed different biologic activity towards heavy metals. It was inhibited by copper, cobalt nickel, iron and lead but was activated by zinc. The GAPDH was purified by ammonium sulphate fractionation and Chelating Sepharose CL-6B chromatography followed by Blue Sepharose CL-6B chromatography. The 150-kDa tetrameric GAPDH showed optimal activity at pH 8.5 and 37°C. The multiple alignment of sequence of the Chironomidae GAPDH with other known species showed 78 - 88% identity to the conserved regions of the GADPH. Bioinformatic analysis unveils substantial N-terminal sequence similarity of GAPDH of Chironomidae larvae to mammalian GADPHs. However, the GADPH of Chironomidae larvae showed different biologic activities and cytotoxicity towards heavy metals. The GAPDH enzyme would undergo adaptive molecular changes through binding at the active site leading to higher tolerance to heavy metals.
authors.affiliation: Department of Pharmacy, Liverpool John Moores University, UK. gurionchong@live.hk
authors.forename: Isaac K W
authors.initials: IK
authors.lastname: Chong
authors.forename: Wing S
authors.initials: WS
authors.lastname: Ho
mesh_terms: D000818:Animals; D002683:Chironomidae / Q000737:chemistry / Q000201:enzymology*; D002846:Chromatography, Affinity; D003300:Copper / Q000737:chemistry / Q000378:metabolism*; D004795:Enzyme Stability; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007814:Larva / Q000737:chemistry / Q000201:enzymology; D020539:Sequence Analysis, Protein; D013696:Temperature; D015032:Zinc / Q000737:chemistry / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D003300:Copper; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D015032:Zinc