Peptide boronic acids Versatile synthetic ligands for affinity chromatography of serine proteinases

David E. Zembower, Cheryl L. Neudauer, Myra J. Wick, Matthew M. Ames

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Peptide boronic acids are potent transition-state analogue inhibitors of serine proteinases. We prepared the peptide boronic acids Ala-Ala-boroPhe (AAbF), targeted at chymotrypsin-like proteinases, and AlaAla-boroVal (AAbV), targeted at elastolytic enzymes. Analogues protected on the N-terminus with the carbonylbenzyloxy (Cbz) group were powerful inhibitors of human neutrophil elastase (HNE) and human cathepsin G (CatG), as well as the non-human counterparts, porcine pancreatic elastase (PPE) and bovine a-chymotrypsin (ChT). Removal of N-Cbz protecting groups and immobilization with Sepharose 6B provided affinity matrices. Columns consisting of the AAbF or AAbV affinity matrix separated a mixture of PPE and ChT. PPE was specifically retained by the AAbV column and ChT was specifically retained by the AAbF column. HNE and CatG were not separated using the AAbF matrix, but were separated with the AAbV matrix. To demonstrate the practical utility of these affinity ligands, HNE was isolated from crude human neutrophil extracts, resulting in an 18-fold purification in one Chromatographie step, with a 41% recovery of elastolytic activity. Because peptide boronic acids can be synthesized having specificity for a wide range of target enzymes, this method is readily adaptable as a general procedure for isolation and purification of serine proteinases.

Original languageEnglish (US)
Pages (from-to)405-413
Number of pages9
JournalInternational Journal of Peptide and Protein Research
Issue number5
StatePublished - 1996


  • Affinity chromatography
  • Boronic acid
  • Cathepsin G
  • Human neutrophil elastase
  • Serine proteinase

ASJC Scopus subject areas

  • Biochemistry


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