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

Journal of Leukocyte Biology

Aravinda Thatte⁵  Martin K. Wild¹  Klaus F. Ley²  Scott Ficarro⁴  Dietmar Vestweber¹  Donald F. Hunt³  Karen R. Snapp⁶ 

[1] Institute of Cell Biology, ZMBE, University of Münster and Max-Planck-Institute of Physiological and Clinical Research, Germany Institute of Cell Biology, ZMBE, University of Münster and Max-Planck-Institute of Physiological and Clinical Research, Germany Institute of Cell Biology, ZMBE, University of Münster and Max-Planck-Institute of Physiological and Clinical Research, Germany;Biomedical Engineering, #Cardiovascular Research Center, and Biomedical Engineering, Biomedical Engineering, #Cardiovascular Research Center, and #Cardiovascular Research Center, and Biomedical Engineering, #Cardiovascular Research Center, andChemistry, ‖Pathology, University of Virginia, Charlottesville; Chemistry, Chemistry, ‖Pathology, University of Virginia, Charlottesville; ‖Pathology, University of Virginia, Charlottesville; Chemistry, ‖Pathology, University of Virginia, Charlottesville;;Chemistry, Chemistry, Chemistry,;Biomedical Engineering, Biomedical Engineering, Biomedical Engineering,;Department of Microbiology and Immunology, Northwestern University, Evanston, Illinois; and Department of Microbiology and Immunology, Northwestern University, Evanston, Illinois; and Department of Microbiology and Immunology, Northwestern University, Evanston, Illinois; and

关键词: arylsulfatase;    selectin;    mass spectrometry;    leukocyte adhesion;   

DOI  :  

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

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 anti-mouse (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.

【 授权许可】

Unknown