Improving binding affinity and stability of peptide ligands by substituting glycines with D-amino acids

Shiyu Chen, David Gfeller, Sergii Buth, Olivier Michielin, Petr Leiman, Christian Heinis

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Improving the binding affinity and/or stability of peptide ligands often requires testing of large numbers of variants to identify beneficial mutations. Herein we propose a type of mutation that promises a high success rate. In a bicyclic peptide inhibitor of the cancer-related protease urokinase-type plasminogen activator (uPA), we observed a glycine residue that has a positive φ{symbol} dihedral angle when bound to the target. We hypothesized that replacing it with a D-amino acid, which favors positive φ{symbol} angles, could enhance the binding affinity and/or proteolytic resistance. Mutation of this specific glycine to D-serine in the bicyclic peptide indeed improved inhibitory activity (1.75-fold) and stability (fourfold). X-ray-structure analysis of the inhibitors in complex with uPA showed that the peptide backbone conformation was conserved. Analysis of known cyclic peptide ligands showed that glycine is one of the most frequent amino acids, and that glycines with positive φ{symbol} angles are found in many protein-bound peptides. These results suggest that the glycine-to-D-amino acid mutagenesis strategy could be broadly applied.

Original languageEnglish (US)
Pages (from-to)1316-1322
Number of pages7
JournalChemBioChem
Volume14
Issue number11
DOIs
StatePublished - Jul 2013
Externally publishedYes

Fingerprint

Glycine
Ligands
Amino Acids
Peptides
Urokinase-Type Plasminogen Activator
Mutation
Cyclic Peptides
Mutagenesis
Dihedral angle
Serine
Conformations
Peptide Hydrolases
X-Rays
X rays
Testing
Neoplasms
Proteins

Keywords

  • Amino acids
  • Bicyclic peptides
  • Glycine
  • Peptides
  • Urokinase

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology

Cite this

Improving binding affinity and stability of peptide ligands by substituting glycines with D-amino acids. / Chen, Shiyu; Gfeller, David; Buth, Sergii; Michielin, Olivier; Leiman, Petr; Heinis, Christian.

In: ChemBioChem, Vol. 14, No. 11, 07.2013, p. 1316-1322.

Research output: Contribution to journalArticle

Chen, Shiyu ; Gfeller, David ; Buth, Sergii ; Michielin, Olivier ; Leiman, Petr ; Heinis, Christian. / Improving binding affinity and stability of peptide ligands by substituting glycines with D-amino acids. In: ChemBioChem. 2013 ; Vol. 14, No. 11. pp. 1316-1322.
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