Abstract
Carbofuran, a potential environmental xenobiotic, has the ability to cross blood brain barrier and to adversely influence brain functions. In the present study, the impact of carbofuran on the biophysical and biochemical properties of rat brain AChE has been evaluatedin vitro. This enzyme was membrane-bound which could be solubilised using Triton-X100 (0.2%, v/v), a nonionic detergent, in the extraction buffer (50 mM phosphate, pH 7.4). The enzyme was highly stable up to one month when stored at-20°C and exhibited optimum activity at pH 7.4 and 37°C. AChE displayed a direct relationship between activity and varying substrate concentrations (acetylthiocholine iodide (ATI)) by following Michaelis-Menten curve. TheKmandVmaxvalues as computed from the Lineweaver-Burk double reciprocal plot of the data were found to be 0.07 mM and 0.066 µmole/mL/min, respectively. The enzyme exhibited IC50value for carbofuran equal to 6.0 nM. The steady-state kinetic studies to determine mode of action of carbofuran on rat brain AChE displayed it to be noncompetitive in nature withKivalue equal to 5 nm. These experiments suggested that rat brain AChE was very sensitive to carbofuran and this enzyme might serve as a significant biomarker of carbofuran induced neurotoxicity.
Highlights
Carbofuran (C12H15NO3; 2,3-dihydro-2,2-dimethyl-7-benzofuranol methylcarbamate, molecular weight, 221.25), commonly known as Furadan, is a broad spectrum carbamate pesticide which is used in various farm practices in order to increase crop productivity
In order to localize the enzyme protein from the rat brain, one part of the tissue was treated with Triton X-100 (0.2%, v/v), a nonionic detergent, in phosphate buffer (50 mM, pH 7.4) to solubilise the membrane-bound protein
Rat brain AChE was found to be membrane-bound which could be solubilised by using Triton X-100, a nonionic detergent
Summary
Carbofuran (C12H15NO3; 2,3-dihydro-2,2-dimethyl-7-benzofuranol methylcarbamate, molecular weight, 221.25), commonly known as Furadan, is a broad spectrum carbamate pesticide which is used in various farm practices in order to increase crop productivity. It has been shown to be generally accumulated in the fat depots and exerts adverse effects on different vital organs such as brain, liver, skeletal muscles, and heart [1, 4, 5]. The neuronal injury of mammalian system due to carbofuran is due to the establishment of oxidative stress [6]. The underlying mechanism of pesticide toxicity relies on generation of oxidative stress [7]. The administration of carbofuran has been found to induce reactive oxygen and nitrogen species in skeletal muscles [8], which are responsible for the peroxidation of membrane phospholipids which disrupt the normal function of lipid bilayer and help develop several pathological conditions
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
