Cholinesterases exhibiting aryl acylamidase activity in human amniotic fluid.
Acetylcholinesterase (EC 3.1.1.7) and butyrylcholinesterase (EC 3.1.1.8) in human amniotic fluid were estimated in the presence of selective inhibitors. Amniotic fluid cholinesterases (mixture of acetylcholinesterase and butyrylcholinesterase) purified by procainamide-Sepharose affinity chromatography exhibited aryl acylamidase activity which was sensitive to serotonin inhibition (a property of aryl acylamidases associated with both acetyl- and butyrylcholinesterases) and tyramine activation (shown exclusively by aryl acylamidase associated with butyrylcholinesterase). Tyramine activation was unaffected in the presence of the selective acetylcholinesterase inhibitor BW284C51 whereas it was abolished in the presence of the selective butyrylcholinesterase inhibitor ethopropazine, suggesting the presence of both types of aryl acylamidases in amniotic fluid, one associated with acetylcholinesterase and the other associated with butyrylcholinesterase. Butyrylcholinesterase and the associated aryl acylamidase activity in the affinity purified enzyme was selectively immunoprecipitated by a polyclonal antibody raised against human serum butyrylcholinesterase. Estimation of the activity ratio of acetylcholinesterase to butyrylcholinesterase in a few samples of amniotic fluid showed that this could vary depending on the butyrylcholinesterase arising from contaminating blood in the samples. Gel electrophoresis under non-denaturing conditions and enzyme staining showed that butyrylcholinesterase band was detectable on the gel in all the samples whereas acetylcholinesterase band was below detectable levels in normal samples but visible in samples from pregnancies of neural tube defect fetuses. It is suggested that the use of selective cholinesterase inhibitors along with gel electrophoresis and immunoprecipitation studies may be useful in the assessment of cholinesterase activities in human amniotic fluid.
Jayanthi
L D
LD
Department of Neurological Sciences, Christian Medical College and Hospital, Vellore, India.
Balasubramanian
N
N
Balasubramanian
A S
AS
D000110:Acetylcholinesterase / Q000302:isolation & purification / Q000378:metabolism*; D000581:Amidohydrolases / Q000378:metabolism*; D000653:Amniotic Fluid / Q000201:enzymology*; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000494:pharmacology; D002091:Butyrylcholinesterase / Q000302:isolation & purification / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007163:Immunosorbent Techniques; D010640:Phenothiazines / Q000494:pharmacology; D011247:Pregnancy; D012701:Serotonin / Q000494:pharmacology; D014439:Tyramine / Q000494:pharmacology
D002800:Cholinesterase Inhibitors; D010640:Phenothiazines; D012701:Serotonin; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase; D000581:Amidohydrolases; C019732:aryl acylamidase; D014439:Tyramine