One-step Bioluminescence ATPase Assay for the Evaluation of Neurotoxic Effects of Metal Ions |
| |
Authors: | Nade?da Nedeljkovi? Anica Horvat |
| |
Institution: | (1) Faculty of Biology, Institute of Physiology and Biochemistry, University of Belgrade, Belgrade, Serbia;(2) Laboratory for Molecular Biology and Endocrinology, “Vinča” Institute of Nuclear Sciences, Belgrade, Serbia |
| |
Abstract: | Summary. Membrane-bound ATPases, such as Na,K-ATPase and nucleotide triphospho-diphosphohydrolase (NTPDase), being one of the first targets of a toxic action are generally considered as good markers for estimating toxicity. A
bioluminescence assay was applied for fast and sensitive evaluation of heavy metals effect on the rat brain synaptosomal membrane
ATPase activity. The assay consists of ATP-consuming reaction catalyzed by synaptic plasma membrane ATPases coupled to the luminescent firefly luciferase reaction, which consumes residual ATP after the course of ATPase reaction. The bioluminescence ATPase assay was applied to study the effect of heavy and transitional metals (Cu2+, Pb2+, Cd2+, Hg2+) on rat brain ATPase activity after assay optimization. All metals applied inhibited synaptic membrane ATPase activity in a concentration dependent manner. The IC
50 values (Hg2+ < Cu2+ < Cd2+ < Pb2+) obtained with the bioluminescence assay were highly correlated with those obtained by the spectrophotometric method. The
fast bioluminescence ATPase assay with small sample and substrate requirements could be adjusted for high-throughput environmental and pharmacological
screening. |
| |
Keywords: | , Bioluminescence ATPase assay, Heavy metal ions, Rat brain, Synaptic plasma membranes, |
本文献已被 SpringerLink 等数据库收录! |
|