Identification of acylpeptide hydrolase as a sensitive site for reaction with organophosphorus compounds and a potential target for cognitive enhancing drugs.
We describe here the purification and identification of a previously unrecognized target for organophosphorus compounds. The target, acylpeptide hydrolase, was isolated as a tritiated-diisopropylfluorophosphate-reactive protein from porcine brain and purified to homogeneity using a combination of ion-exchange and gel-filtration chromatography. Biochemical characterization and internal sequence analysis confirmed identity. Acylpeptide hydrolase was found to be potently inhibited by the organophosphorus compounds chlorpyrifosmethyl oxon, dichlorvos, and diisopropylfluorophosphate (20-min IC(50) values of 18.3 +/- 2.0, 118.7 +/- 9.7, and 22.5 +/- 1.2 nM, respectively). The in vitro sensitivity of acylpeptide hydrolase toward these compounds is between six and ten times greater than that of acetylcholinesterase (AChE), making it a target of pharmacological and toxicological significance. We show that, in vivo, acylpeptide hydrolase is significantly more sensitive than AChE to inhibition by dichlorvos and that the inhibition is more prolonged after a single dose of inhibitor. Furthermore, using dichlorvos as a progressive inhibitor, it was possible to show that acylpeptide hydrolase is the only enzyme in the brain capable of hydrolyzing the substrate N-acetyl-alanyl-p-nitroanilide. A concentration of 154 +/- 27 pmol of acylpeptide hydrolase/gram of fresh rat brain was also deduced by specific labeling with tritiated-diisopropylfluorophosphate. We also suggest that, by comparison of structure-activity relationships, acylpeptide hydrolase may be the target for the cognitive-enhancing effects of certain organophosphorus compounds. Acylpeptide hydrolase cleaves N(alpha)-acylated amino acids from small peptides and may be involved in regulation of neuropeptide turnover, which provides a new and plausible mechanism for its proposed cognitive enhancement effect.
Richards
P G
PG
Medical Research Council Toxicology Unit, University of Leicester, Leicester, United Kingdom. pgr3@le.ac.uk
Johnson
M K
MK
Ray
D E
DE
D000818:Animals; D001921:Brain / Q000201:enzymology; D002219:Carbamates / Q000378:metabolism / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000378:metabolism / Q000494:pharmacology; D003071:Cognition / Q000187:drug effects*; D004006:Dichlorvos / Q000494:pharmacology; D006868:Hydrolysis; D008970:Molecular Weight; D018697:Nootropic Agents / Q000494:pharmacology*; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D010447:Peptide Hydrolases / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D000085822:Prolyl Oligopeptidases; D051381:Rats; D020539:Sequence Analysis, Protein; D012697:Serine Endopeptidases / Q000378:metabolism; D013552:Swine; D000521:alpha-MSH / Q000378:metabolism
D002219:Carbamates; D002800:Cholinesterase Inhibitors; D018697:Nootropic Agents; D009943:Organophosphorus Compounds; D000521:alpha-MSH; D004006:Dichlorvos; D010447:Peptide Hydrolases; C038220:acylaminoacyl-peptidase; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases