Branched-chain-amino-acid aminotransferase assay using radioisotopes

Biochim Biophys Acta. 1989 Sep 15;992(3):397-9. doi: 10.1016/0304-4165(89)90103-7.

Abstract

A method for the assay of the activity of branched-chain-amino-acid aminotransferase from Escherichia coli has been developed. Radioactive isoleucine is used, the radioactive oxoacid formed in the enzymic reaction is converted to its p-nitrophenylhydrazone, and the hydrazone is extracted into toluene based scintillation liquid. The small reaction tubes containing the toluene layer and the reaction mixture as a water layer are placed into liquid scintillation vials and counted for radioactivity. The radioactive amino acid remaining in the water layer causes only a rather low background.

MeSH terms

  • Carbon Radioisotopes
  • Escherichia coli / enzymology*
  • Kinetics
  • Radioisotope Dilution Technique
  • Scintillation Counting
  • Transaminases / analysis*
  • Transaminases / metabolism

Substances

  • Carbon Radioisotopes
  • Transaminases
  • branched-chain-amino-acid transaminase