Abundant tissue butyrylcholinesterase and its possible function in the acetylcholinesterase knockout mouse.
We have described recently an acetylcholinesterase (AChE) knockout mouse. While comparing the tissue distribution of AChE and butyrylcholinesterase (BChE), we found that extraction buffers containing Triton X-100 strongly inhibited mouse BChE activity. In contrast, buffers with Tween 20 caused no inhibition of BChE. Conventional techniques grossly underestimated BChE activity by up to 15-fold. In Tween 20 buffer, the intestine, serum, lung, liver, and heart had higher BChE than AChE activity. Only brain had higher AChE than BChE activity in AChE +/+ mice. These findings contradict the dogma, based mainly on observations in Triton X-100 extracts, that BChE is a minor cholinesterase in animal tissues. AChE +/- mice had 50% of normal AChE activity and AChE -/- mice had none, but all mice had similar levels of BChE activity. BChE was inhibited by Triton X-100 in all species tested, except rat and chicken. Inhibition was reversible and competitive with substrate binding. The active site of rat BChE was unique, having an arginine in place of leucine at position 286 (human BChE numbering) in the acyl-binding pocket of the active site, thus explaining the lack of inhibition of rat BChE by Triton X-100. The generally high levels of BChE activity in tissues, including the motor endplate, and the observation that mice live without AChE, suggest that BChE has an essential function in nullizygous mice and probably in wild-type mice as well.
Li
B
B
Eppley Institute, University of Nebraska Medical Center, Omaha 68198-6805, USA.
Stribley
J A
JA
Ticu
A
A
Xie
W
W
Schopfer
L M
LM
Hammond
P
P
Brimijoin
S
S
Hinrichs
S H
SH
Lockridge
O
O
D000110:Acetylcholinesterase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002091:Butyrylcholinesterase / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009045:Motor Endplate / Q000201:enzymology*; D018482:Muscle, Skeletal / Q000201:enzymology*; D009928:Organ Specificity; D011409:Propylene Glycols / Q000494:pharmacology; D051381:Rats; D013045:Species Specificity; D013770:Tetraisopropylpyrophosphamide / Q000494:pharmacology
D002800:Cholinesterase Inhibitors; D011409:Propylene Glycols; D013770:Tetraisopropylpyrophosphamide; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase