TY - JOUR
T1 - Cholinergic elements in a human choriocarcinoma cell line
AU - Fant, Michael
AU - Speeg, Kermit V.
AU - Harper, Sandra
AU - Harbison, Raymond D.
N1 - Funding Information:
* Supported, in part, by USPHS Grants ES00782, ES00267 and GM15431 and the Veterans Administration Medical Research Program. t Present address: Children’s Hospital Medical Center, Boston, MA, U.S.A. $ M. Fant is the recipient of a Vivian Allen M.D./Ph.D. Fellowship. § Author to whom all correspondence should be addressed: Dr. Raymond D. Harbison, Department of Pharmacology, Vanderbilt Medical Center, Nashville, TN 37232, U.S.A. ‘1C . H. Smith, personal communication.
PY - 1981/5/1
Y1 - 1981/5/1
N2 - Components of the cholinergic system have been identified in the JEG choriocarcinoma cell line. [3H]Quinuclidinyl benzilate (QNB) was used to identify high affinity muscarinic binding sites in a whole cell preparation. Specific binding was saturable with respect to QNB concentration and revealed a binding site density of 27 fmoles/mg protein. The bimolecular rates of association, 2.24 × 107 M-1 min-1, and dissociation, 4.2 × 10-3min-1, revealed a dissociation constant (Kd) of 180 pM which agreed closely with that derived from saturation isotherms, 245 pM. Muscarinic antagonists and agonists were able to compete effectively for these binding sites, whereas non-muscarinic compounds were not. Cholinesterase activity was also demonstrated with substrate preference consistent with that of acetyl-cholinesterase (acetylcholine > acetyl-β-methylcholine > butyrylcholine) hydrolyzing 2.42 ± 0.19 × 10-3 μmoles acetylcholine·min-1 ·(mg protein)-1. No choline acetyltransferase activity was detected in these cells, however.
AB - Components of the cholinergic system have been identified in the JEG choriocarcinoma cell line. [3H]Quinuclidinyl benzilate (QNB) was used to identify high affinity muscarinic binding sites in a whole cell preparation. Specific binding was saturable with respect to QNB concentration and revealed a binding site density of 27 fmoles/mg protein. The bimolecular rates of association, 2.24 × 107 M-1 min-1, and dissociation, 4.2 × 10-3min-1, revealed a dissociation constant (Kd) of 180 pM which agreed closely with that derived from saturation isotherms, 245 pM. Muscarinic antagonists and agonists were able to compete effectively for these binding sites, whereas non-muscarinic compounds were not. Cholinesterase activity was also demonstrated with substrate preference consistent with that of acetyl-cholinesterase (acetylcholine > acetyl-β-methylcholine > butyrylcholine) hydrolyzing 2.42 ± 0.19 × 10-3 μmoles acetylcholine·min-1 ·(mg protein)-1. No choline acetyltransferase activity was detected in these cells, however.
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U2 - 10.1016/0006-2952(81)90041-1
DO - 10.1016/0006-2952(81)90041-1
M3 - Article
C2 - 7195203
AN - SCOPUS:0019423805
VL - 30
SP - 967
EP - 970
JO - Biochemical Pharmacology
JF - Biochemical Pharmacology
SN - 0006-2952
IS - 9
ER -