Recognition of protein-linked glycans as a determinant of peptidase activity

Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):E679-E688. doi: 10.1073/pnas.1615141114. Epub 2017 Jan 17.


The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. This insight builds upon the paradigm of how peptidases recognize substrates and provides a molecular understanding of glycoprotein degradation.

Keywords: O-glycopeptidase; O-glycosylation; glycoprotein; metallopeptidase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Escherichia coli / genetics
  • Fetuins / metabolism
  • Glycopeptides / metabolism
  • Glycosylation
  • Mucins / metabolism
  • Peptide Hydrolases / chemistry
  • Peptide Hydrolases / genetics
  • Peptide Hydrolases / metabolism*
  • Polysaccharides / metabolism*
  • Protein Conformation
  • Protein Processing, Post-Translational


  • Fetuins
  • Glycopeptides
  • Mucins
  • Polysaccharides
  • Peptide Hydrolases

Associated data

  • PDB/5KD2
  • PDB/5KD5
  • PDB/5KD8
  • PDB/5KDJ
  • PDB/5KDN
  • PDB/5KDS
  • PDB/5KDU
  • PDB/5KDV
  • PDB/5KDW
  • PDB/5KDX