Abstract
A cDNA for Drosophila choline acetyltransferase (ChAT) was expressed in E. coli and the recombinant enzyme partially purified. Kinetic analysis yielded the following constants for the recombinant enzyme; KmAcCoA = 29 μM, KmCoA = 25 μM, Kmcholine = 330 μM and Kmacetylcholine = 2 mM. The recombinant Drosophila enzyme, like the enzyme from other species, exhibited an increase in activity as a function of increased salt concentration. Chemical modification studies using dithio-bis-nitro-2-carboxylate, butanedione, and diethylpyrocarbonate showed that the recombinant enzyme contains active site cysteine, arginine, and histidine residues. These studies demonstrate that the recombinant Drosophila ChAT possesses the same catalytic properties as the enzyme from a variety of other sources.
Original language | English |
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Pages (from-to) | 119-124 |
Number of pages | 6 |
Journal | Brain Research Bulletin |
Volume | 24 |
Issue number | 1 |
DOIs | |
State | Published - Jan 1990 |
Bibliographical note
Funding Information:This research was supported in part by grants AG 05893 and AG 08013 from the National Institute on Aging.
Keywords
- Choline acetyltransferase (ChAT)
- Drosophila
- Kinetics
- Recombinant enzyme
ASJC Scopus subject areas
- General Neuroscience