Formation of multiple complexes between beta-dystroglycan and dystrophin family products.
Beta-dystroglycan is expressed in a wide variety of tissues and has generally been reported with an Mr of 43 kDa, sometimes accompanied with a 31 kDa protein assumed to be a truncated product. This molecule was recently identified as the anomalous beta-dystroglycan expressed in various carcinoma cell lines. We produced and characterized a G5 polyclonal antibody specific to beta-dystroglycan that is directed against the C-terminal portion of the molecule. We provide evidence that beta-dystroglycan may vary in size and properties by studying different Xenopus tissues. Besides normal beta-dystroglycan with an Mr of 43 kDa in smooth and cardiac muscle and sciatic nerve extracts, we found it in skeletal muscle and brain proteins with an Mr of 38 and 65 kDa, respectively. Glycosylation properties and proteolytic susceptibilities of these different beta-dystroglycans are analysed and compared in this work. Crosslinking experiments with various beta-dystroglycan preparations obtained from skeletal and cardiac muscles and brain gave rise to specific new covalent products with Mr of 125 kDa (doublet band), or 120 and 130 kDa, or 140 and 240 kDa, respectively. We provide evidence, using various similar beta-dystroglycan preparations, that the immunoprecipitation procedure with G5 specific polyclonal antibody allows consistent pelleting of various dystrophin-family isoforms. Skeletal muscles from Xenopus reveals the presence of two distinct beta-dystroglycan complexes, one with dystrophin and another one which involves alpha-dystrobrevin. Cardiac muscle and brain from Xenopus are shown to contain three beta-dystroglycan complexes related to various dystrophin-family isoforms. Dystrophin or alpha-dystrobrevin or Dp71 were found in cardiac muscle and dystrophin or Dp180 or Up71 in brain. This variability in the relationship between beta-dystroglycan and dystrophin-family isoforms suggests that each protein--currently known as dystrophin associated protein--could not be present in each of these complexes.
Royuela
M
M
Department of Cell Biology and Genetics, University of Alcalá, E-28871 Alcalá de Henares, Madrid, Spain.
Chazalette
D
D
Hugon
G
G
Paniagua
R
R
Guerlavais
V
V
Fehrentz
J A
JA
Martinez
J
J
Labbe
J P
JP
Rivier
F
F
Mornet
D
D
D000818:Animals; D001921:Brain / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D049030:Dystroglycans; D016189:Dystrophin / Q000378:metabolism*; D049029:Dystrophin-Associated Proteins; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism*; D009130:Muscle, Smooth / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D012584:Sciatic Nerve / Q000378:metabolism; D014982:Xenopus laevis
D003598:Cytoskeletal Proteins; D016189:Dystrophin; D049029:Dystrophin-Associated Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D020033:Protein Isoforms; C098885:dystrobrevin; D049030:Dystroglycans