Force spectroscopy between acetylcholinesterase molecule and its natural substrate to study the effects of inhibitors and reactivators on enzyme activity.
The force spectrum (FS) between acetylcholinesterase (AChE) molecule and its natural substrates acetylcholine (ACh) and the influences of AChE inhibitors and reactivators have been investigated with atomic force microscopy (AFM) at single molecule level in real-time. AChE and ACh were covalently immobilized onto the surfaces of gold-plated mica and Si3N4 tip of the atomic force microscope respectively. First, AChE was imaged in image mode of AFM and one of AChE molecules was selected as the center of the scanning. Then scanning mode was changed into force scanning mode and FS was recorded in a frequency of 5 x s(-1). Solutions of drugs or toxicants can be injected from the fluid-in tube of the fluid cell at any desired time. The FS between ideally immobilized normal AChE, Inhibited AChE or aged AChE and ACh each had their own shape features. The influences of drugs or toxicants on these features could be observed in real-time on the screen of the computer. These results demonstrated that AFM force spectroscopy could be used as a new method to study the effects of drugs and toxicants on the activity of the enzyme in pharmacology and toxicology.
Yingge
Z
Z
Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing, PRC. zhangyingge@hotmail.com
Delu
Z
Z
Chunli
B
B
Chen
W
W
D000109:Acetylcholine / Q000737:chemistry / Q000378:metabolism*; D000110:Acetylcholinesterase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002619:Chemical Warfare Agents / Q000737:chemistry / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D002801:Cholinesterase Reactivators / Q000737:chemistry / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000737:chemistry; D007700:Kinetics; D018625:Microscopy, Atomic Force; D010091:Oximes; D011220:Pralidoxime Compounds / Q000737:chemistry / Q000494:pharmacology; D011726:Pyridinium Compounds / Q000737:chemistry / Q000494:pharmacology; D014101:Torpedo
D002619:Chemical Warfare Agents; D002800:Cholinesterase Inhibitors; D002801:Cholinesterase Reactivators; D004800:Enzymes, Immobilized; D010091:Oximes; D011220:Pralidoxime Compounds; D011726:Pyridinium Compounds; D000110:Acetylcholinesterase; C022870:asoxime chloride; D000109:Acetylcholine; C028797:pralidoxime