Paper Report for: Hsiao_2004_J.Agric.Food.Chem_52_5340
Reference
Title: Purification and characterization of acetylcholinesterase from oriental fruit fly [Bactrocera dorsalis (Hendel)] (Diptera: Tephritidae) Hsiao YM, Lai JY, Liao HY, Feng HT Ref: Journal of Agricultural and Food Chemistry, 52:5340, 2004 : PubMed
An acetylcholinesterase (AChE, EC 3.1.1.7) was purified from the head of the insecticide susceptible oriental fruit fly, Bactrocera dorsalis (Hendel), by affinity chromatography of Triton X-100 extract. The degree of purification was about 8183-fold with recoveries of 52%. The molecular mass of purified AChE was 116 kDa for its native protein (nonreduced form) and 61 kDa for its subunits (reduced form) as revealed on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), suggesting that the homodimer of AChE linked with disulfide bonds. Nondenaturing PAGE of the purified AChE revealed only one molecular form. The maximum velocities (V(max)) for hydrolyzing acetylthiocholine (ATC), propionylthiocholine, and S-butyrylthiocholine were 833.3, 222.2, and 57.5 micromol/min/mg, and the Michaelis constants (K(m)) were 87.9, 26.9, and 195.3 microM, respectively. More than 97% of AChE activity was inhibited by 10 microM eserine or BW284C51, but only 53% of the activity was inhibited by ethopropazine at the same concentration. On the basis of the substrate and inhibitor specificities, the purified enzyme appeared to be a true AChE. Nevertheless, the purified AChE exhibited some distinctive characteristics including (i) a lack of the substrate inhibition phenomenon when using ATC as the hydrolyzing substrate and (ii) a higher V(m) value for ATC than AChE from other insect species. These biochemical properties may show that AChE purified from the oriental fruit fly may have structural differences from those of other insect species.
        
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Hsiao YM, Lai JY, Liao HY, Feng HT (2004) Purification and characterization of acetylcholinesterase from oriental fruit fly [Bactrocera dorsalis (Hendel)] (Diptera: Tephritidae) Journal of Agricultural and Food Chemistry52: 5340-6
Hsiao YM, Lai JY, Liao HY, Feng HT (2004) Journal of Agricultural and Food Chemistry52: 5340-6