Preparation of antibodies specific to choline acetyltransferase from bovine caudate nucleus and immunohistochemical localization of the enzyme

C. Cozzari, B. K. Hartman

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

Choline acetyltransferase (CATase; acetylCoA:choline O-acetyltransferase, EC 2.3.1.6) has been purified from bovine caudate nucleus. The specific activity of the pure enzyme was 120-160 μmol of acetylcholine formed per mg per min. The purified enzyme separated into two bands (band A and band B) when electrophoresed in a pH 4.3 gradient gel. Both bands exhibited CATase activity. Antisera were prepared in rabbits to each form. Immunotitrations using either antisera resulted in quantitative precipitation of CATase activity from enzyme preparations at all stages of purification. Both antisera produced single immunoprecipitin lines in double-diffusion experiments when run against the enzyme at each stage of purity. Immunoprecipitin lines cut from double-diffusion gels contained CATase activity, demonstrating that the observed reaction was due to antibody interaction with the enzyme. Immunoelectrophoresis also showed a single immunoprecipitin line against the purified enzyme as well as against a crude caudate extract. These results indicated that immunochemically pure and specific antisera have been prepared to bovine CATase and that both the A and B forms of the enzyme have common antigenic sites. The antisera were utilized to localize the enzyme in bovine brain. The localization was exclusively neuronal with both antisera, and both forms of CATase were present in the same population of neurons. Subtle differences in the precise intracellular distribution of enzyme were observed with the two antisera, indicating that although the two molecular forms of CATase have antigenic sites in common they are not antigenically identical.

Original languageEnglish (US)
Pages (from-to)7453-7457
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume77
Issue number12 II
DOIs
StatePublished - 1980

Fingerprint Dive into the research topics of 'Preparation of antibodies specific to choline acetyltransferase from bovine caudate nucleus and immunohistochemical localization of the enzyme'. Together they form a unique fingerprint.

  • Cite this