title: Laminin-binding protein 120 from brain is closely related to the dystrophin-associated glycoprotein, dystroglycan, and binds with high affinity to the major heparin binding domain of laminin.
abstract: When brain proteins separated by SDS-polyacrylamide gel electrophoresis (PAGE) and transferred to nitrocellulose are probe with 125I-labeled laminin, a single broad band of approximately 120 kDa binds laminin specifically. We show here by two-dimensional electrophoresis and protein microsequencing that this band consists of two distinct laminin-binding proteins. One of these is the amyloid precursor protein. The other, laminin-binding protein (LBP) 120, is closely related to the dystrophin-associated glycoprotein, dystroglycan (156 kDa); 5 peptides from purified bovine brain LBP120, ranging in size from 7 to 19 residues, are up to 100% identical to the predicted amino acid sequence of muscle dystroglycan (ibraghimov-Beskrovanaya, O., Ervasti, J. M., Leveille, C. J., Slaughter, C. A., Sernett, S. W., and Campbell, K. P. (1992) Nature 355, 696-702). These protein microsequence data support the data of Ibraghimov-Beskrovnaya et al., which suggest that the dystroglycan precursor is processed into 120/156- and 43-kDa proteins. Moreover, the data suggest a revision in the position of the proposed cleavage site of the precursor. The glycosylation and extracellular localization of LBP120/dystroglycan are consistent with it being a cell surface laminin receptor. LBP120/dystroglycan, either as a native protein, or following SDS-PAGE and transfer to nitrocellulose, binds with high affinity (Kd = 90 nM) to a proteolytic fragment of laminin (E3) containing the major heparin binding domain. This binding is Ca(2+)-dependent and inhibited by low concentrations of heparin. Thus, LBP120/dystroglycan is a major non-integrin laminin receptor whose high affinity interaction with laminin may reflect a structural role in brain and muscle.
authors.affiliation: Centre for Research in Neurosciences, Montreal General Hospital Research Institute, McGill University, Montreal, Canada.
authors.forename: S H
authors.initials: SH
authors.lastname: Gee
authors.forename: R W
authors.initials: RW
authors.lastname: Blacher
authors.forename: P J
authors.initials: PJ
authors.lastname: Douville
authors.forename: P R
authors.initials: PR
authors.lastname: Provost
authors.forename: P D
authors.initials: PD
authors.lastname: Yurchenco
authors.forename: S
authors.initials: S
authors.lastname: Carbonetto
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002642:Chick Embryo; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D049030:Dystroglycans; D016189:Dystrophin / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D017474:Receptors, Laminin / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D003598:Cytoskeletal Proteins; D016189:Dystrophin; D007797:Laminin; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D017474:Receptors, Laminin; D049030:Dystroglycans; D006493:Heparin