N-glycosylation of Siglec-15 decreases its lysosome-dependent degradation and promotes its transportation to the cell membrane

Biochem Biophys Res Commun. 2020 Nov 26;533(1):77-82. doi: 10.1016/j.bbrc.2020.08.111. Epub 2020 Sep 11.

Abstract

Siglec-15 was recently reported to be an immunosuppressive molecule that is expressed by tumor-associated macrophages and upregulated in some solid tumors. Targeting Siglec-15 is a potential strategy for normalization cancer immunotherapy. Here, we identified the important post-translational modification, N-glycosylation of Siglec-15, which is regulated by glucose uptake. Using a series of glycosidase and glycosylation inhibitors, we demonstrated that Siglec-15 was completely N-glycosylated in vitro and in vivo. The precise glycosylation site was determined. N-glycosylation stabilized Siglec-15 by decreasing its lysosome-dependent degradation. Siglec-15 subcellular distribution detected by immunofluorescence indicated that N-glycosylation promoted Siglec-15 transportation to the cell membrane. The collective observations indicate that targeting the N-glycosylation of Siglec-15 may be an effective supplement to immunotherapy.

Keywords: Lysosome-dependent degradation; N-Glycosylation; Siglec-15; Subcellular distribution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Membrane / metabolism
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Immunoglobulins / metabolism*
  • Lysosomes / metabolism*
  • Membrane Proteins / metabolism*
  • Protein Transport
  • Proteolysis

Substances

  • Immunoglobulins
  • Membrane Proteins
  • SIGLEC15 protein, human