A Peptide Mimetic of 5-Acetylneuraminic Acid-Galactose Binds with High Avidity to Siglecs and NKG2D

PLoS One. 2015 Jun 25;10(6):e0130532. doi: 10.1371/journal.pone.0130532. eCollection 2015.

Abstract

We previously identified several peptide sequences that mimicked the terminal sugars of complex glycans. Using plant lectins as analogs of lectin-type cell-surface receptors, a tetravalent form of a peptide with the sequence NPSHPLSG, designated svH1C, bound with high avidity to lectins specific for glycans with terminal 5-acetylneuraminic acid (Neu5Ac)-galactose (Gal)/N-acetylgalactosamine (GalNAc) sequences. In this report, we show by circular dichroism and NMR spectra that svH1C lacks an ordered structure and thus interacts with binding sites from a flexible conformation. The peptide binds with high avidity to several recombinant human siglec receptors that bind preferentially to Neu5Ac(α2,3)Gal, Neu5Ac(α2,6)GalNAc or Neu5Ac(α2,8)Neu5Ac ligands. In addition, the peptide bound the receptor NKG2D, which contains a lectin-like domain that binds Neu5Ac(α2,3)Gal. The peptide bound to these receptors with a KD in the range of 0.6 to 1 μM. Binding to these receptors was inhibited by the glycoprotein fetuin, which contains multiple glycans that terminate in Neu5Ac(α2,3)Gal or Neu5Ac(α2,6)Gal, and by sialyllactose. Binding of svH1C was not detected with CLEC9a, CLEC10a or DC-SIGN, which are lectin-type receptors specific for other sugars. Incubation of neuraminidase-treated human peripheral blood mononuclear cells with svH1C resulted in binding of the peptide to a subset of the CD14+ monocyte population. Tyrosine phosphorylation of siglecs decreased dramatically when peripheral blood mononuclear cells were treated with 100 nM svH1C. Subcutaneous, alternate-day injections of svH1C into mice induced several-fold increases in populations of several types of immune cells in the peritoneal cavity. These results support the conclusion that svH1C mimics Neu5Ac-containing sequences and interacts with cell-surface receptors with avidities sufficient to induce biological responses at low concentrations. The attenuation of inhibitory receptors suggests that svH1C has characteristics of a checkpoint inhibitor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Galactose / metabolism*
  • Humans
  • Leukocytes, Mononuclear / drug effects
  • Mice
  • N-Acetylneuraminic Acid / chemistry
  • N-Acetylneuraminic Acid / metabolism*
  • NK Cell Lectin-Like Receptor Subfamily K / metabolism*
  • Peptides / chemistry
  • Peptides / metabolism
  • Plant Lectins / chemistry
  • Plant Lectins / metabolism*
  • Sialic Acid Binding Immunoglobulin-like Lectins / chemistry
  • Sialic Acid Binding Immunoglobulin-like Lectins / metabolism

Substances

  • Klrk1 protein, mouse
  • NK Cell Lectin-Like Receptor Subfamily K
  • Peptides
  • Plant Lectins
  • Sialic Acid Binding Immunoglobulin-like Lectins
  • N-Acetylneuraminic Acid
  • Galactose

Grants and funding

The work was supported by Susavion Biosciences, Inc. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.