Acetylcholinesterases from the Disease Vectors Aedes aegypti and Anopheles gambiae: Functional Characterization and Comparisons with Vertebrate Orthologues

Mosquitoes of the Anopheles (An.) and Aedes (Ae.) genus are principal vectors of human diseases including malaria, dengue and yellow fever. Insecticide-based vector control is an established and important way of preventing transmission of such infections. Currently used insecticides can efficiently control mosquito populations, but there are growing concerns about emerging resistance, off-target toxicity and their ability to alter ecosystems. A potential target for the development of insecticides with reduced off-target toxicity is the cholinergic enzyme acetylcholinesterase (AChE). Herein, we report cloning, baculoviral expression and functional characterization of the wild-type AChE genes (ace-1) from An. gambiae and Ae. aegypti, including a naturally occurring insecticide-resistant (G119S) mutant of An. gambiae. Using enzymatic digestion and liquid chromatography-tandem mass spectrometry we found that the secreted proteins were post-translationally modified. The Michaelis-Menten constants and turnover numbers of the mosquito enzymes were lower than those of the orthologous AChEs from Mus musculus and Homo sapiens. We also found that the G119S substitution reduced the turnover rate of substrates and the potency of selected covalent inhibitors. Furthermore, non-covalent inhibitors were less sensitive to the G119S substitution and differentiate the mosquito enzymes from corresponding vertebrate enzymes. Our findings indicate that it may be possible to develop selective non-covalent inhibitors that effectively target both the wild-type and insecticide resistant mutants of mosquito AChE.     

Effects of arsenate and copper on the algal communities in polluted lakes in the northern parts of Sweden assayed by PICT (Pollution-Induced Community Tolerance)

The tolerance for arsenate and copper in the carbon dioxide fixation activity of phytoplankton communities coming from lakes around the smelter at Rönnskär at the Swedish east-coast were measured during three years (1989–1991). The smelter have for several decades discharged arsenic and heavy metals into the air, and their concentrations in the lakes were clearly correlated with the distance from the smelter. The tolerance of communities from the three most polluted lakes were higher than communities from reference lakes with background concentrations of arsenic and copper. In accordance with the PICT concept it is indicated that those communities have been affected by these substances. These communities also had lower diversity than the others, but no clear correlation could be done with phytoplankton species number, or between phytoplankton biomass and pollution levels.