Albumin is a major protein component of transverse tubule vesicles isolated from skeletal muscle.
Despite multiple procedures used to isolate transverse tubule vesicles from rabbit skeletal muscle, few proteins have been identified and shown to be specific to transverse tubule vesicles. Markers for purified transverse tubules have included high affinity dihydropyridine binding, cholesterol content, Mg2+-ATPase activity, (Na+,K+)-ATPase activity, and [3H] ouabain binding. Despite these markers, few proteins from purified transverse tubules can be unequivocally identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In this report we have biochemically and immunologically identified rabbit albumin as a major component of purified transverse tubule membranes from rabbit skeletal muscle. Albumin composed between 5.1 and 9.8% (n = 4) of the total protein in purified transverse tubules based on scans of SDS-PAGE. Furthermore, albumin and other serum proteins are present in preparations of transverse tubules and triads but not in light sarcoplasmic reticulum. Extraction of triads with low concentrations of saponin or sodium dodecyl sulfate completely removes albumin without removing intrinsic membrane proteins. Our results suggest that albumin and other serum proteins are present in the lumen of preparations of transverse tubules and albumin may be used as a marker for the transverse tubules when analyzed on SDS gels.
Knudson
C M
CM
Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City 52242.
Campbell
K P
KP
D000418:Albumins / Q000032:analysis*; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002501:Centrifugation, Zonal / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D008870:Microtubules / Q000032:analysis* / Q000648:ultrastructure; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis* / Q000648:ultrastructure; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000032:analysis* / Q000648:ultrastructure
D000418:Albumins