Dystrophin-glycoprotein complex is highly enriched in isolated skeletal muscle sarcolemma.
mAbs specific for protein components of the surface membrane of rabbit skeletal muscle have been used as markers in the isolation and characterization of skeletal muscle sarcolemma membranes. Highly purified sarcolemma membranes from rabbit skeletal muscle were isolated from a crude surface membrane preparation by wheat germ agglutination. Immunoblot analysis of subcellular fractions from skeletal muscle revealed that dystrophin and its associated glycoproteins of 156 and 50 kD are greatly enriched in purified sarcolemma vesicles. The purified sarcolemma was also enriched in novel sarcolemma markers (SL45, SL/TS230) and Na+/K(+)-ATPase, whereas t-tubule markers (alpha 1 and alpha 2 subunits of dihydropyridine receptor, TS28) and sarcoplasmic reticulum markers (Ca2(+)-ATPase, ryanodine receptor) were greatly diminished in this preparation. Analysis of isolated sarcolemma by SDS-PAGE and densitometric scanning demonstrated that dystrophin made up 2% of the total protein in the rabbit sarcolemma preparation. Therefore, our results demonstrate that although dystrophin is a minor muscle protein it is a major constituent of the sarcolemma membrane in skeletal muscle. Thus the absence of dystrophin in Duchenne muscular dystrophy may result in a major disruption of the cytoskeletal network underlying the sarcolemma in dystrophic muscle.
Ohlendieck
K
K
Howard Hughes Medical Institute, University of Iowa College of Medicine, Iowa City 52242.
Ervasti
J M
JM
Snook
J B
JB
Campbell
K P
KP
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient; D016189:Dystrophin / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis*; D015151:Immunoblotting; D046911:Macromolecular Substances; D009124:Muscle Proteins / Q000032:analysis*; D011817:Rabbits; D012508:Sarcolemma / Q000737:chemistry*; D013347:Subcellular Fractions / Q000737:chemistry; D014909:Wheat Germ Agglutinins
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D016189:Dystrophin; D006023:Glycoproteins; D046911:Macromolecular Substances; D009124:Muscle Proteins; D014909:Wheat Germ Agglutinins