Porin-type 1 proteins in sarcoplasmic reticulum and plasmalemma of striated muscle fibres.
The location of porin-type 1 proteins in mammalian striated muscle has been assessed using immunogold electron microscopy with an anti-porin 31HL monoclonal antibody as the primary antibody. Gold particles were found on the mitochondrial outer membrane, the sarcoplasmic reticulum and plasmalemma in longitudinal sections of rat and rabbit skeletal muscle and rabbit and sheep cardiac muscle. The relative densities of gold particles in the mitochondrial outer membrane, sarcoplasmic reticulum and plasmalemma were 7:3:1 in white sternomastoid muscle, for example. Skeletal and cardiac sarcoplasmic reticulum vesicles, which had been fractionated by discontinuous sucrose density centrifugation, were subjected to SDS-polyacrylamide gel electrophoresis and Western blotting. The anti-porin 31HL monoclonal antibody detected a band of relative molecular mass (M(r)) 31,000 in all muscle sarcoplasmic reticulum vesicle fractions and also in liver mitochondria. The intensity of immunostaining of the sarcoplasmic reticulum fractions was 2.5-10% that of mitochondrial outer membranes per microgram of membrane protein blotted. Contamination of the sarcoplasmic reticulum fractions by mitochondrial outer membrane was < 0.75% as determined from the specific activity of monoamine oxidase. Thus, only a small part of the porin detected in sarcoplasmic reticulum vesicles can be attributed to mitochondrial contamination. These results show that porin-type1 immunoreactivity is not restricted to mitochondria but found in the sarcoplasmic reticulum and plasmalemma of both mammalian skeletal and cardiac muscle.
Junankar
P R
PR
Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra, Australia.
Dulhunty
A F
AF
Curtis
S M
SM
Pace
S M
SM
Thinnes
F P
FP
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry*; D002467:Cell Nucleus / Q000737:chemistry; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000737:chemistry; D016253:Microscopy, Immunoelectron; D008930:Mitochondria, Liver / Q000737:chemistry; D008970:Molecular Weight; D018656:Muscle Fibers, Fast-Twitch / Q000737:chemistry; D018657:Muscle Fibers, Slow-Twitch / Q000737:chemistry; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000648:ultrastructure; D009206:Myocardium / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000276:immunology; D018272:Porins / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012519:Sarcoplasmic Reticulum / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D010455:Peptides; D018272:Porins