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A SIMPLIFIED RADIOCHEMICAL ASSAY FOR CHOLINE ACETYLTRANSFERASE

A SIMPLIFIED RADIOCHEMICAL ASSAY FOR CHOLINE ACETYLTRANSFERASE Departmeat of Pharmacology Tufts University School of Medicine 136 Harrison Avenue Boston, Massachusetts 02111 (Received 3 January 1967) CHOLINE acetyltransferase (acetyl-CoA:choline 0-acetyltransferase, I.U.B. 2.3.1.6) is an enzyme of major interest in neurochemistry. Until recently, studies with this enzyme were limited by the availability of the bioassays, described by CHANG and GADDUM (1933) and the much less sensitive colorimetric method of HESTRIN (1949). SCHUBERTH (1963, 1966) developed a radiochemical assay for the enzyme, but the method required an acetyl-CoA generating system since it did not distinguish acetylCoA from ACh. Selective precipitation methods were developed by MCCAMAN and HUNT(1965) and ALPERT, KISLIUK SHUSTER and (1966), in which ACh is precipitated as the reineckate, and by FONNUM (1966), in which the tetraphenylborate salt is formed. These precipitation methods are specific and their sensitivity is limited only by the available specific activity of the substrate, but the tedium of washing and centrifuging the precipitated product remains. A gas chromatographic method designed by STAVINOHA RYAN (1965) is somewhat cumbersome for several and concurrent assays or for use in enzyme purification studies. We have designed and evaluated an assay method for choline acetyltransferase which is rapid and precise. It was originally developed for use http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neurochemistry Wiley

A SIMPLIFIED RADIOCHEMICAL ASSAY FOR CHOLINE ACETYLTRANSFERASE

Schrier, B. K.; Snuster, L.
Journal of Neurochemistry , Volume 14 (10) – Oct 1, 1967
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Publisher
Wiley
Copyright
Copyright © 1967 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0022-3042
eISSN
1471-4159
DOI
10.1111/j.1471-4159.1967.tb09509.x
Publisher site
See Article on Publisher Site

Abstract

Departmeat of Pharmacology Tufts University School of Medicine 136 Harrison Avenue Boston, Massachusetts 02111 (Received 3 January 1967) CHOLINE acetyltransferase (acetyl-CoA:choline 0-acetyltransferase, I.U.B. 2.3.1.6) is an enzyme of major interest in neurochemistry. Until recently, studies with this enzyme were limited by the availability of the bioassays, described by CHANG and GADDUM (1933) and the much less sensitive colorimetric method of HESTRIN (1949). SCHUBERTH (1963, 1966) developed a radiochemical assay for the enzyme, but the method required an acetyl-CoA generating system since it did not distinguish acetylCoA from ACh. Selective precipitation methods were developed by MCCAMAN and HUNT(1965) and ALPERT, KISLIUK SHUSTER and (1966), in which ACh is precipitated as the reineckate, and by FONNUM (1966), in which the tetraphenylborate salt is formed. These precipitation methods are specific and their sensitivity is limited only by the available specific activity of the substrate, but the tedium of washing and centrifuging the precipitated product remains. A gas chromatographic method designed by STAVINOHA RYAN (1965) is somewhat cumbersome for several and concurrent assays or for use in enzyme purification studies. We have designed and evaluated an assay method for choline acetyltransferase which is rapid and precise. It was originally developed for use

Journal

Journal of Neurochemistry – Wiley

Published: Oct 1, 1967

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