Isolation and characterization of a laminin-binding protein from rat and chick muscle.
A major laminin-binding protein (LBP), distinct from previously described LBPs, has been isolated from chick and rat skeletal muscle (Mr 56,000 and 66,000, respectively). The purified LBPs from the two species were shown to be related antigenically and to have similar NH2-terminal amino acid sequences and total amino acid compositions. Protein blots using laminin and laminin fragments provided evidence that this LBP interacts with the major heparin-binding domain, E3, of laminin. Studies on the association of this LBP with muscle membrane fractions and reconstituted lipid vesicles indicate that this protein can interact with lipid bilayers and has properties of a peripheral, not an integral membrane protein. These properties are consistent with its amino acid sequence, determined from cDNAs (Clegg et al., 1988). Examination by light and electron microscopy of the LBP antigen distribution in skeletal muscle indicated that the protein is localized primarily extracellularly, near the extracellular matrix and myotube plasmalemma. While a form of this LBP has been identified in heart muscle, it is present at low or undetectable levels in other tissues examined by immunocytochemistry indicating that it is probably a muscle-specific protein. As this protein is localized extracellularly and can bind to both membranes and laminin, it may mediate myotube interactions with the extracellular matrix.
Howard Hughes Medical Institute, University of California, San Francisco 94143-0724.
D E
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Hall
K A
KA
Frazer
B C
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Hann
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Reichardt
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D002135:Calcium-Binding Proteins; D002155:Calsequestrin; D002352:Carrier Proteins / Q000032:analysis / Q000302:isolation & purification*; D002462:Cell Membrane / Q000032:analysis; D002499:Centrifugation, Density Gradient; D002645:Chickens; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000032:analysis; D005260:Female; D007118:Immunoassay; D007150:Immunohistochemistry; D007797:Laminin / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis* / Q000648:ultrastructure; D011817:Rabbits; D051381:Rats
D002135:Calcium-Binding Proteins; D002155:Calsequestrin; D002352:Carrier Proteins; D007797:Laminin; D008565:Membrane Proteins; C057137:Casq1 protein, rat