Properties of chicken cardiac dystrophin.
We investigated the presence of dystrophin by immunoblot and immunofluorescence analyses, negative staining, rotatory shadowing and immunogold electron microscopy in chicken cardiac muscle. Saponin was found to be better than Triton X-100 for providing a new 'dystrophin-enriched' solution for use in biochemical studies of the molecule. By Western blot analysis, only a 400-kDa band was revealed with polyclonal antibodies directed against a central region (residues 1178-1723) of the dystrophin molecule and no cross-reactions with other proteins or degraded products were observed. Specific cleavage of the dystrophin molecule showed that the central rod-shaped domain corresponded to a resistant 'core'. This structure might rigidify the protein. By immunofluorescence, dystrophin was localized at the periphery of cardiac ventricular cells. The molecule was examined by electron microscopy and found to have variable lengths (140-160 nm for the monomeric from and about 260 +/- 10 nm or more for oligomeric forms). These oligomeric structures are considered to be associated molecules which are only partially overlapped lengthwise. The precise distribution of dystrophin within the cardiac muscle was determined by visualisation of gold particles in immuno-electron microscopy. Gold particles were found on the sarcolemma with no evidence of any association with cytoplasmic structures. The present data provide further details on the cardiac dystrophin molecule and suggest that its capacity of self-association may elasticize the dystrophin dimer.
Fabbrizio
E
E
Pathologie Moléculaire du Muscle, INSERM U300, Faculté de Pharmacie, Montpellier, France.
Harricane
M C
MC
Pons
F
F
Leger
J
J
Mornet
D
D
D000818:Animals; D002645:Chickens; D016189:Dystrophin / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D010450:Endopeptidases; D005455:Fluorescent Antibody Technique; D006868:Hydrolysis; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D009206:Myocardium / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
D016189:Dystrophin; D010450:Endopeptidases