BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Modification of Carboxyl Residues of Proteins with Pyridoxamine as a Fluorophore

  • Kwon, Oh-Shin (Department of Biochemistry, College of Natural Sciences, Kyungpook National University)
  • Received : 1996.01.10
  • Published : 1996.05.31
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Abstract

A general procedure to quantitate the reaction of carbodiimides with carboxy groups of proteins is described. Pyridoxamine reacts with the o-acylisourea intermediate generated during the reaction of carboxyl residues with carbodiimides. The extent of the reaction is determined by measuring the spectroscopic properties, absorption and emission, of pyridoxyl residues covalently attached to the proteins. Resolved pig brain aspartate aminotransferase (apoenzyme), inactivated by 1-ethyl-3-(3-dimethylamino propyl) carbodiimide, reacts with $[^{3}H]pyridoxamine$. After trypsin digestion, one peptide labeled with radioactive pyridoxyl was separated by reverse phase HPLC.

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