Table II.
Increase in the specific activity of NAAT-III by affinity chromatography
| Binding Fraction | Non-Bound Fraction | |||||||
|---|---|---|---|---|---|---|---|---|
| Ligand | NA | α-KG | α-KG | A-2-C | NA | α-KG | α-KG | A-2-C |
| Solute of specific elution buffer | NA | NA | α-KG | A-2-C | NA | NA | α-KG | A-2-C |
| pkat mg−1 protein | ||||||||
| Before affinity chromatography | 519 | 261 | ||||||
| After affinity chromatography | 96 | 599 | 706 | 575 | 1298 | 601 | 926 | 868 |
Binding and non-bound fractions of DEAE were applied to affinity chromatography columns with three ligands and eluted by four specific elution buffers. For the ligands, NA was used as a substrate, 2-oxoglutarate (α-KG) as an amino group acceptor, and azetidine-2-carboxylic acid (A-2-C) as a substrate analog. The concentration of the solute in the specific elution buffers was 1 mm.