Identification and characterization of ligands for L-selectin in the kidney. I. Versican, a large chondroitin sulfate proteoglycan, is a ligand for L-selectin.
Ligands for a leukocyte adhesion molecule, L-selectin, are expressed not only in the specific vascular endothelium in lymph nodes and Peyer's patches but also in the extravascular tissues such as the brain white matter, choroid plexus and the kidney distal straight tubuli. However, the biological significance of these extravascular ligands is currently unknown. We now report the purification and characterization of a novel extravascular ligand for L-selectin in the kidney using a tubule-derived cell line, ACHN. Binding of L-selectin-IgG chimera (LEC-IgG) to the isolated ligand was specifically blocked with either (i) anti-L-selectin mAb, (ii) EDTA, (iii) fucoidan, (iv) chondroitin sulfate (CS) B or CS E, or (v) treatment with chondroitinases. Partial amino acid sequencing, Western blotting and immunoprecipitation analyses showed that a major ligand for L-selectin in ACHN cells is versican of 1600 kDa. Histochemical as well as biochemical analyses verified that a versican subspecies in the kidney was indeed reactive with L-selectin. Studies with cell lines including those derived from the kidney indicated that a certain glycoform and/or splice form of versican is reactive with L-selectin. Under pathological conditions such as those induced by unilateral ureteral obstruction, versican was shed from the distal straight tubuli and became localized in the adjacent vascular bundles around which a substantial leukocyte infiltration was concomitantly observed. Possible involvement of versican in leukocyte trafficking into the kidney under diseased conditions is discussed.
Kawashima
H
H
Department of Bioregulation, Biomedical Research Center, Osaka University Medical School, Suita, Japan.
Li
Y F
YF
Watanabe
N
N
Hirose
J
J
Hirose
M
M
Miyasaka
M
M
D000818:Animals; D015153:Blotting, Western; D011508:Chondroitin Sulfate Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007684:Kidney Tubules / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D019041:L-Selectin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D037181:Lectins, C-Type; D008297:Male; D010449:Peptide Mapping; D011233:Precipitin Tests; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000378:metabolism; D017421:Sequence Analysis; D014407:Tumor Cells, Cultured; D053675:Versicans
D011508:Chondroitin Sulfate Proteoglycans; D037181:Lectins, C-Type; D011994:Recombinant Proteins; C508322:VCAN protein, human; C508324:Vcan protein, rat; D019041:L-Selectin; D053675:Versicans