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 [3H]pyridoxamine. After trypsin digestion, one peptide labeled with radioactive pyridoxyl was separated by reverse phase HPLC.
| Original language | English |
|---|---|
| Pages (from-to) | 215-220 |
| Number of pages | 6 |
| Journal | Journal of Biochemistry and Molecular Biology |
| Volume | 29 |
| Issue number | 3 |
| State | Published - 1996 |
Keywords
- Aspartate aminotransferase
- Carbodiimide
- Pyridoxamine
- Spectroscopic properties