Expression, Purification, and Characterization of Recombinant Drosophila Choline Acetyltransferase

Donghai Wu, Norbert Schormann, Wei Lian, Johann Deisenhofer, Louis B. Hersh

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


Abstract: A cDNA for Drosophila choline acetyltransferase (EC; ChAT) was fused with a polyhistidine sequence and expressed in Escherichia coli. The recombinant enzyme was purified to a specific activity of 500 μmol/min/mg of protein using metal affinity chromatography and ion exchange chromatography. Kinetic properties of the recombinant enzyme did not differ significantly from those previously determined. Circular dichroism (CD) spectra revealed that the secondary structure of the enzyme is largely μ‐helical. Intrinsic fluorescence spectra of the enzyme indicate that its tryptophan residues are buried. Neither CD nor fluorescence spectra changed significantly in the presence of substrates. The cysteine content of the recombinant Drosophila ChAT was determined to be 16 in the absence and 22 in the presence of 6 M guanidine hydrochloride. Finally, crystallization of recombinant Drosophila ChAT was achieved.

Original languageEnglish
Pages (from-to)1416-1422
Number of pages7
JournalJournal of Neurochemistry
Issue number4
StatePublished - Oct 1993


  • Drosophila choline acetyltransferase
  • Enzyme crystallization
  • Enzyme physicochemical characterization
  • Recombinant enzyme

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience


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