Hericium erinaceus isolectins recognize mucin-type O-glycans as tumor-associated carbohydrate antigens on the surface of K562 human leukemia cells

Int J Biol Macromol. 2018 Dec;120(Pt A):1093-1102. doi: 10.1016/j.ijbiomac.2018.08.115. Epub 2018 Aug 24.

Abstract

In Hericium erinaceus mushroom fruiting body, two different lectin groups, HEL1 and HEL2, were identified by using peptide mass fingerprinting based on customized protein sequence databases derived from RNA-Seq data. The HEL2 group included four isoforms designated HEL2a-d. Codon-optimized genes encoding HEL1, HEL2a, and HEL2b were expressed in Escherichia coli to produce fully active soluble proteins designated rHEL1, rHEL2a, and rHEL2b. Interestingly, these lectins showed different molecular weights: approximately 15 kDa for rHEL1 and approximately 120 kDa for rHEL2a and rHEL2b under non-denaturing conditions. rHEL2a and rHEL2b exhibited agglutination activities, but rHEL1 did not show any agglutination activity toward animal erythrocytes. The hemagglutination activity of rHEL2 lectins was strongly inhibited by glycoproteins containing mucin-type O-glycans. Glycan array analysis and isothermal titration calorimetry revealed that rHEL2 isolectins interacted strongly with O-glycans harboring the core 1 O-glycan motif, Galβ(1,3)GalNAc. Moreover, the glycan binding specificities of rHEL2 isolectins were comparable to that of peanut agglutinin in their ability to recognize O-glycans attached to leukosialin as tumor-associated carbohydrate antigens on the surface of K562 human leukemia cells. These results indicate that rHEL2 isolectins could be used as a powerful tool for analyzing mucin-type O-glycans expressed on the surface of cancer cells.

Keywords: Cancer biology; Cancer cell surface; Core 1 O-linked glycan; Diagnosis assay; Extended core 1 O-glycan; Hericium erinaceus; Human leukemia cell line; Isolectin; K562; Tumor-associated carbohydrate antigens.

MeSH terms

  • Antigens, Tumor-Associated, Carbohydrate / chemistry*
  • Antigens, Tumor-Associated, Carbohydrate / immunology
  • Antigens, Tumor-Associated, Carbohydrate / isolation & purification
  • Basidiomycota / chemistry*
  • Basidiomycota / genetics
  • Basidiomycota / immunology
  • Carbohydrate Sequence / genetics
  • Humans
  • K562 Cells
  • Lectins / chemistry
  • Lectins / genetics
  • Lectins / immunology*
  • Leukemia / genetics
  • Leukemia / immunology*
  • Leukemia / pathology
  • Mucins / chemistry
  • Mucins / genetics
  • Mucins / immunology
  • Polysaccharides / chemistry
  • Polysaccharides / genetics
  • Polysaccharides / immunology
  • Protein Binding

Substances

  • Antigens, Tumor-Associated, Carbohydrate
  • Lectins
  • Mucins
  • Polysaccharides