Biochemical and toxicological properties of two acetylcholinesterases from the common bed bug, Cimex lectularius.
We examined the molecular and enzymatic properties of two acetylcholinesterases (AChEs; ClAChE1 and ClAChE2) from the common bed bug, Cimex lectularius. Native polyacrylamide gel electrophoresis followed by activity staining and Western blotting revealed that ClAChE1 is the main catalytic enzyme and is abundantly expressed in various tissues. Both ClAChEs existed in dimeric form connected by a disulfide bridge and were attached to the membrane via a glycophosphatidylinositol anchor. To determine their kinetic and inhibitory properties, both ClAChE1 and ClAChE2 were in vitro expressed in Sf9 cells using a baculovirus expression system. ClAChE1 showed higher catalytic efficiency toward acetylcholine, supporting the hypothesis that ClAChE1 plays a major role in postsynaptic transmission. An inhibition assay revealed that ClAChE1 is generally more sensitive to organophosphates and carbamates examined although ClAChE2 was >4000-fold more sensitive to malaoxon than ClAChE1. The relatively higher correlation between the in vitro ClAChE1 inhibition and the in vivo toxicity suggested that ClAChE1 is the more relevant toxicological target for organophosphates and carbamates. Although the physiological function of ClAChE2 remains to be elucidated, ClAChE2 also appears to have neuronal functions, as judged by its tissue distribution and molecular and kinetic properties. Our findings help expand our knowledge on insect AChEs and their toxicological properties.
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea.
Chae Eun
CE
Hwang
Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-742, Republic of Korea.
Young Ho
YH
Kim
Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-742, Republic of Korea.
Deok Ho
DH
Kwon
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea.
Keon Mook
KM
Seong
Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-742, Republic of Korea.
Jae Young
JY
Choi
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea; Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-742, Republic of Korea.
Yeon Ho
YH
Je
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea; Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-742, Republic of Korea. Electronic address: shlee22@snu.ac.kr.
Si Hyeock
SH
Lee
D000005:Abdomen; D000109:Acetylcholine / Q000378:metabolism; D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D001511:Bedbugs / Q000187:drug effects / Q000201:enzymology*; D001921:Brain / Q000201:enzymology; D005121:Extremities; D006257:Head; D019476:Insect Proteins / Q000378:metabolism*; D007306:Insecticides / Q000633:toxicity; D012469:Salivary Glands / Q000201:enzymology; D013909:Thorax / Q000201:enzymology
D019476:Insect Proteins; D007306:Insecticides; D000110:Acetylcholinesterase; D000109:Acetylcholine
Acetylcholinesterase; Carbamate; Cimex lectularius; Kinetics; Molecular form; Organophosphate