A hematopoietic cell L-selectin ligand that is distinct from PSGL-1 and displays N-glycan-dependent binding activity

Blood. 2000 Oct 15;96(8):2765-74.


Human hematopoietic progenitor cells express L-selectin and also express PSGL-1, a ligand for all selectins. Using a shear-based adhesion assay, a hematopoietic cell L-selectin ligand (HCLL) that is expressed on the hematopoietic cell line KG1a and on normal human hematopoietic progenitors was previously identified. To characterize the structural biology of HCLL and to define its relationship to PSGL-1, the effects of chemical and enzymatic treatments on HCLL activity of KG1a cells and membrane preparations were analyzed. Protease digestions and chemical treatments of KG1a cells and membranes indicated that HCLL is an integral membrane glycoprotein. Glycosidase digestions of membrane protein preparations and metabolic treatments of KG1a cells with glycosylation processing modifiers revealed that L-selectin binding determinants on HCLL are sialofucosylated structures presented on complex-type N-glycans. Adhesion assays and biochemical studies showed that this glycoprotein is also expressed on circulating blasts in native acute leukemias. HCLL is distinguishable from PSGL-1: (1) KG1a cells sorted for PSGL-1 expression had equivalent HCLL activity; (2) anti-PSGL-1 blocking antibodies and proteases known to eliminate L-selectin binding to PSGL-1 had no effect on HCLL binding activity of KG1a cells; (3) blasts from native leukemias with low expression of PSGL-1 and CD34 display high HCLL activity; and (4) despite high level expression of PSGL-1, HCLL activity was absent on HL60 cells. These data provide first evidence of a naturally expressed membrane L-selectin ligand expressing binding determinant(s) on an N-linked glycoconjugate. This novel ligand may help mediate L-selectin-dependent cell-cell adhesive interactions within the cytoarchitecture of the bone marrow microenvironment. (Blood. 2000;96:2765-2774)

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acute Disease
  • Bromelains / pharmacology
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Fucose / analysis
  • Glycoside Hydrolases / pharmacology
  • Glycosylation
  • HL-60 Cells / chemistry
  • Hematopoietic Stem Cells / metabolism*
  • Humans
  • L-Selectin / metabolism*
  • Leukemia, Myeloid / pathology
  • Ligands
  • Membrane Glycoproteins / analysis
  • Membrane Glycoproteins / isolation & purification*
  • Membrane Glycoproteins / metabolism
  • Molecular Sequence Data
  • Neoplasm Proteins / isolation & purification
  • Neoplasm Proteins / metabolism
  • Neoplastic Stem Cells / chemistry
  • Neuraminidase / pharmacology
  • Polysaccharides / physiology*
  • Sialic Acids / analysis
  • Tumor Cells, Cultured / chemistry


  • Ligands
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • P-selectin ligand protein
  • Polysaccharides
  • Sialic Acids
  • L-Selectin
  • Fucose
  • Bromelains
  • Glycoside Hydrolases
  • Neuraminidase