Lectin and epidermal growth factor domains of P-selectin at physiologic density are the recognition unit for leukocyte binding

R. M. Gibson, G. S. Kansas, T. F. Tedder, B. Furie, B. C. Furie*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

P-selectin is an integral membrane glycoprotein on stimulated platelets and endothelial cells that serves as a receptor for leukocytes. To estimate the density of P-selectin in membranes necessary to support adhesion, we incorporated purified P-selectin at varying concentrations into phospholipid bilayers that encapsulated glass microspheres. Maximal binding of these lipospheres to HL60 cells, a P-selectin ligand-expressing cell line, was approached at a P-selectin density of about 100 molecules per μm2; half- maximal binding was observed at about 50 to 60 molecules per μm2. Compatible results were obtained with P-selectin expressed on Chinese hamster ovary cells. The P-selectin density on stimulated platelets was estimated to be 150 to 200 molecules/μm2. To identify the domains of P-selectin required for HL60 cell binding, chimeras of P-selectin and L-selectin were stably expressed in Chinese hamster ovary cells and clones that expressed the chimeras at the estimated physiologic density were selected. Chimeras containing the P-selectin lectin and epidermal growth factor (EGF) domains or the lectin, EGF, and short consensus repeats bound HL60 cells equivalently, but a chimera containing the P-selectin lectin domain alone bound HL60 cells much less well. These results indicate that at a physiologically relevant P- selectin density on membrane surfaces, the lectin, and EGF domains of P- selectin are together required for optimal leukocyte binding.

Original languageEnglish (US)
Pages (from-to)151-158
Number of pages8
JournalBlood
Volume85
Issue number1
DOIs
StatePublished - Jan 1 1995

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Fingerprint Dive into the research topics of 'Lectin and epidermal growth factor domains of P-selectin at physiologic density are the recognition unit for leukocyte binding'. Together they form a unique fingerprint.

Cite this