Binding of function-blocking mAbs to mouse and human P-selectin glycoprotein ligand-1 peptides with and without tyrosine sulfation

Aravinda Thatte, Scott Ficarro, Karen R. Snapp, Martin K. Wild, Dietmar Vestweber, Donald F. Hunt, Klaus F. Ley

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

P-selectin glycoprotein ligand-1 (PSGL-1) mediates rolling of leukocytes on P-selectin-expressing endothelial cells under shear flow. Function-blocking monoclonal antibodies (mAbs) against mouse and human PSGL-1 recognize an anionic segment at the N-terminus of PSGL-1. High affinity interaction of PSGL-1 with P-selectin requires sulfation of tyrosines 46, 48, and 51 (human) or 54 and 56 (mouse). We tested binding of two anti-human (KPL1 and PL1) and two antimouse (4RA10 and 2PH1) PSGL-1 mAbs to synthetic peptides of N-terminus of human and mouse PSGL-1 and found binding to be independent of tyrosine sulfation. In peptide-blocking experiments, sulfated and nonsulfated human and mouse peptides competed with antibody binding to PSGL-1 expressed on myeloid cells. Arylsulfatase treatment significantly reduced P-selectin binding but had no effect on antibody binding. Our data show, in three independent assay systems, that function-blocking antibodies to mouse or human PSGL-1 do not require sulfation of N-terminal tyrosines for binding.

Original languageEnglish (US)
Pages (from-to)470-477
Number of pages8
JournalJournal of Leukocyte Biology
Volume72
Issue number3
StatePublished - Sep 1 2002
Externally publishedYes

Keywords

  • Arylsulfatase
  • Leukocyte adhesion
  • Mass spectrometry
  • Selectin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Cell Biology

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