Isolation and partial characterization of high affinity laminin receptors in neural cells.
Neural cells in culture (NG-108, PC12, chick dorsal root ganglion, chick spinal cord, and rat astrocytes) bind laminin with an apparent Kd of congruent to 10(-9) M. Laminin affinity chromatography of chick brain membranes washed with 150 mM NaCl and eluted with 0.2 M glycine buffer, pH 3.5, yields a single protein with an apparent molecular mass of 67 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Isoelectric focusing and peptide mapping indicate that the 67-kDa protein is distinct from bovine serum albumin (68 kDa) but indistinguishable from high affinity laminin receptors isolated from skeletal muscle. After electroblotting onto nitrocellulose paper and probing with 125I-laminin, this putative laminin receptor binds laminin specifically (100 ng/ml). A second protein (congruent to 120-140 kDa) is also detected with 125I-laminin (100 ng/ml) in the laminin affinity-purified membrane proteins. Both 67- and congruent to 120-140-kDa proteins can be laminin affinity-purified from cultures enriched for neurons (greater than 90%) following metabolic labeling with [35S]methionine. Our data suggest that neural cells (dorsal root ganglion, central nervous system neurons, astrocytes, and several neural cell lines) have high affinity binding sites for laminin and that two membrane proteins, 67- and congruent to 120-140-kDa, are responsible at least in part for this binding.
Douville
P J
PJ
Centre for Research in Neuroscience, Montreal General Hospital Research Institute, McGill University, Quebec, Canada.
Harvey
W J
WJ
Carbonetto
S
S
D000818:Animals; D001253:Astrocytes / Q000276:immunology; D001921:Brain / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D002478:Cells, Cultured; D002645:Chickens; D002846:Chromatography, Affinity; D005455:Fluorescent Antibody Technique; D005727:Ganglia, Spinal / Q000276:immunology; D007797:Laminin / Q000378:metabolism*; D008970:Molecular Weight; D009474:Neurons / Q000276:immunology*; D051381:Rats; D011971:Receptors, Immunologic / Q000302:isolation & purification* / Q000378:metabolism; D017474:Receptors, Laminin; D013116:Spinal Cord / Q000276:immunology
D007797:Laminin; D011971:Receptors, Immunologic; D017474:Receptors, Laminin