Identification and characterization of ligands for L-selectin in the kidney. III. Characterization of L-selectin reactive heparan sulfate proteoglycans.
L-Selectin, a leukocyte adhesion molecule, mediates leukocyte rolling on the endothelium and plays a critical role in leukocyte recruitment at inflammatory sites as well as in lymphocyte homing. We have previously shown that L-selectin reactive chondroitin sulfate and heparan sulfate proteoglycans (HSPGs) are both expressed in the distal tubules of the kidney and that versican is one of the chondroitin sulfate-type ligands. In the present study, we characterized the heparan sulfate-type ligand(s) in more detail. The molecular sizes of HSPGs were approximately 600 kDa with core protein sizes of 160 and 180 kDa. Western blotting analysis showed that L-selectin reactive HSPGs were neither agrin nor perlecan, major basement membrane HSPGs in the kidney. The binding to L-selectin was mediated by the lectin domain of L-selectin in a Ca2+-dependent manner and required heparan sulfate side chains, but not sialic acid. To our knowledge, this is the first biochemical characterization of the L-selectin reactive heparan sulfate proteoglycan(s) in the distal tubules of the kidney.
Watanabe
N
N
Department of Bioregulation, Biomedical Research Center, Osaka University Medical School, Suita, 565-0871, Japan.
Kawashima
H
H
Li
Y F
YF
Miyasaka
M
M
D000818:Animals; D001665:Binding Sites; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D066298:In Vitro Techniques; D007686:Kidney Tubules, Distal / Q000378:metabolism*; D019041:L-Selectin / Q000378:metabolism*; D008024:Ligands; D008297:Male; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar
D019812:Heparan Sulfate Proteoglycans; D008024:Ligands; D019041:L-Selectin