Correlation of binding sites for diisopropyl phosphorofluoridate with cholinesterase and neuropathy target esterase in membrane and cytosol preparations from hen.
To find new putative target(s) for organophosphorus induced delayed neurotoxicity (OPIDN), we investigated the biochemical and pharmacological characteristics of [3H] diisopropyl phosphorofluoridate (DFP) binding to membrane and cytosol preparations from the brain and spinal cord of hens. Specific [3H]DFP binding was determined by subtracting non-specific binding from total binding. The binding sites of [3H]DFP, an organophosphate that induces OPIDN, were found not only on membrane but also in cytosol. Reduction of subsequent ex vivo specific [3H]DFP binding by in vivo pretreatment with unlabeled DFP was found in cytosol, not membrane. The reduced binding lasted to the onset of OPIDN, especially in spinal cord. These results suggest that the specific DFP binding sites in cytosol, rather than on membrane, are the most important with regard to the initiation of OPIDN. Inhibitors of cholinesterase (ChE) and neuropathy target esterase (NTE) other than DFP did not affect specific [3H]DFP binding to either membranes or cytosol in vivo. Additionally, inhibition of the activities of these esterases by these compounds was not consistent with either the degree of inhibition of the [3H]DFP binding or a time-dependent manner of OPIDN. These results suggest that DFP binding site(s) involved in the initiation of OPIDN may be different from the active sites of ChE and NTE.
Kamata
R
R
Department of Veterinary Pharmacology, Faculty of Agriculture, Iwate University, Morioka, Japan. u9996001@iwate-u.ac.jp
Saito
S
S
Suzuki
T
T
Takewaki
T
T
Kobayashi
H
H
D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors*; D002645:Chickens / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000378:metabolism* / Q000633:toxicity*; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000633:toxicity*; D005260:Female; D007531:Isoflurophate / Q000378:metabolism* / Q000633:toxicity; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism
D002800:Cholinesterase Inhibitors; D004791:Enzyme Inhibitors; D007531:Isoflurophate; D002265:Carboxylic Ester Hydrolases; C021859:neurotoxic esterase