Palladium enolates derived from β-ketocarbonyl compounds are stable in air and moisture. This stability was utilized to develop a peptide purification system using β-ketoamide as a small affinity tag in aqueous media. A solid-supported palladium complex efficiently captured and released β-ketoamide-tagged molecules, allowing for the purification and identification of ligand-binding sites in proteins.
Palladium enolates derived from & beta;-ketocarbonyl compounds serve as key intermediates in various catalytic asymmetric reactions. We found that the palladium enolate formed from & beta;-ketoamide is stable in air and moisture and we applied this property to develop a peptide purification system using & beta;-ketoamide as a small affinity tag in aqueous media. A solid-supported palladium complex successfully captured & beta;-ketoamide-tagged molecules as palladium enolates and released them in high yield upon acid treatment. Optimum conditions for the catch and release of tagged peptides from a mixture of untagged peptides were established. To demonstrate the value of this methodology in identifying the binding site of a ligand to its target protein, we purified and identified a peptide containing the ligand-binding site from the tryptic digest of cathepsin B labelled with a covalent cathepsin B inhibitor containing a & beta;-ketoamide tag.
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