Cytochemical, histological, and phylogenetic distribution of a 38,000-dalton protein associated with transverse tubules.
A major protein in detergent extracts of skeletal muscle appears at 38,000 daltons in electrophoretic separations. Previous investigations have provided indirect evidence that a 38-kD skeletal muscle protein is membrane associated, and this inference has served as the basis for speculations on 38-kD protein function. In the present study, affinity purified, polyclonal antisera against 38-kD protein from skeletal muscle are produced for immunolocalization and biochemical assays. Immunoblots of two-dimensional electrophoretic separations show that this protein is heterogenously charged at pI approximately 6.4. This 38-kD protein is not extracted from muscle in low ionic strength or high ionic strength buffers, in isotonic buffers from pH 4 to pH 8 or in buffers containing 5 mM EGTA. The 38-kD protein is extracted, however, by isotonic, pH 7.0 buffer containing 1.0% Triton-X. Light microscope observations using indirect immunofluorescence of anti-38-kD labeled tissue show the protein distributed in a reticular pattern within cross-sectional muscle but not at the cell surface. Longitudinal sections show the protein concentrated in periodic, transverse bands. Purified fractions of muscle plasma membrane analyzed by immunoblotting contain 38-kD protein. Immunoblots using anti-38 kD show that this protein is present in all vertebrate skeletal muscle examined, however, the protein is present only in cardiac muscle that contains transverse tubules. The antibody does not recognize aldolase, another 38-kD striated muscle protein.
Tidball
J G
JG
Division of Biomedical Sciences, University of California, Riverside 92521-0121.
Gadus
M V
MV
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D002645:Chickens; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis; D015151:Immunoblotting; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis*; D009206:Myocardium / Q000032:analysis; D010802:Phylogeny; D011817:Rabbits; D011894:Rana pipiens
D009124:Muscle Proteins; D005634:Fructose-Bisphosphate Aldolase