Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin

PLoS One. 2015 Jun 26;10(6):e0129608. doi: 10.1371/journal.pone.0129608. eCollection 2015.

Abstract

O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. Research in this field, however, is hampered by the lack of suitable probes to identify, accumulate, and purify the O-GlcNAcylated proteins. We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. Study on AAL2 may enable us to design a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylglucosamine / chemistry*
  • Acetylglucosamine / metabolism
  • Agrocybe / metabolism*
  • Amino Acid Sequence
  • Binding Sites
  • Glycosylation
  • Lectins / chemistry*
  • Lectins / metabolism
  • Metals / chemistry
  • Metals / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism
  • Protein Binding
  • Protein Conformation
  • Sequence Alignment
  • Structure-Activity Relationship
  • Substrate Specificity
  • Surface Plasmon Resonance

Substances

  • Lectins
  • Metals
  • Polysaccharides
  • Acetylglucosamine

Associated data

  • PDB/4TQJ
  • PDB/4TQK
  • PDB/4TQM

Grants and funding

This work was supported by the funding from Chinese Ministry of Science and Technology 973 Programs (2011CB911100, 2011CB910304) to DCW, National Natural Science Foundation of China (31370849), the Chinese 111 Project (B06018), and from Wuhan University (2042014kf0226) to HS.