Inactivation of γaminobutyric acid aminotransferase by various amine buffers

Mark Hans Hopkins*, Katherine A. Bichler, Ting Su, Cheryl L. Chamberlain, Richard B. Silverman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

It is hypothesized that buffers capable of forming a Schiff base with the PLP of y-aminobutyric acid aminotransferase (GABA-AT) may lead to denaturation and inactivation of the enzyme. On the basis of this hypothesis three new methods for the selective destruction of GABA-AT in GABAse (a commercial bacterial source of a mixture of GABA-AT and succinic semialdehyde dehydrogenase [SSDH]) and from pig brain are described: (1) dialysis against a primary or secondary amine buffer; (2) gel filtration with a primary or secondary amine buffer as eluent; (3) inactivation with gabaculine followed by dialysis or gel filtration with pyrophosphate buffer. The SSDH activity in GABAse, which remains unchanged by all of these methods, may then be used in a coupled assay to measure the activity of GABA-AT from different sources. These results also suggest that the use of primary and secondary amine buffers should be avoided when inhibitors are being tested with GABA-AT.

Original languageEnglish (US)
Pages (from-to)195-199
Number of pages5
JournalJournal of Enzyme Inhibition and Medicinal Chemistry
Volume6
Issue number3
DOIs
StatePublished - 1992

Keywords

  • Amine buffers
  • Bis-tris
  • Bis-tris propane
  • Pyrophosphate
  • Succinic semialdehyde dehydrogenase
  • Tris
  • γAminobutyric acid aminotransferase

ASJC Scopus subject areas

  • Pharmacology
  • Drug Discovery

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