Characterization of a 30-kDa peripheral nerve glycoprotein that binds laminin and heparin.
We have shown previously that a bovine peripheral nerve protein with a molecular mass of about 30 kDa binds laminin in blot overlay assay. In this paper, we have characterized this 30-kDa laminin-binding protein (LBP30). LBP30 was extracted from the crude bovine peripheral nerve membranes at pH 12 or by 0.5 M NaCl but not by 2% Triton X-100. LBP30 bound to heparin-Sepharose in the presence of 0.5 M NaCl. The results of lectin staining indicated that LBP30 contained both terminally sialylated and nonsialylated Ser/Thr-linked oligosaccharides. LBP30 bound laminin-2 as well as laminin-1 but not fibronectin or collagen type IV. When immobilized LBP30 was incubated with the crude peripheral nerve membrane extracts, all of the endogenous peripheral nerve laminin chain isoforms, the alpha1, alpha2, beta1, beta2, and gamma1 chains, were detected bound to LBP30. The binding of LBP30 to laminin was inhibited by heparin, heparan sulfate, dextran sulfate, or NaCl but was not affected significantly by chondroitin sulfate, dextran, or EDTA. Although LBP30 bound to laminin-1 denatured with SDS in a nonreducing condition, the binding was reduced drastically when laminin-1 was denatured with SDS in a reducing condition, suggesting that the binding of LBP30 is somewhat dependent on the high order structure of laminin-1. Immunohistochemical analysis demonstrated the broad distribution of LBP30 in the perineurium and endoneurium of bovine peripheral nerve. These results indicate that LBP30 is a laminin- and heparin-binding glycoprotein localized in the perineurium and endoneurium of bovine peripheral nerve.
Saito
F
F
Department of Neurology and Neuroscience, Teikyo University School of Medicine, Tokyo 173, Japan.
Yamada
H
H
Sunada
Y
Y
Hori
H
H
Shimizu
T
T
Matsumura
K
K
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*; D017933:Peripheral Nervous System / Q000378:metabolism*; D011485:Protein Binding
D006023:Glycoproteins; D007797:Laminin; D009419:Nerve Tissue Proteins; D006493:Heparin