title: Proteomic analysis of the dysferlin protein complex unveils its importance for sarcolemmal maintenance and integrity.
abstract: Dysferlin is critical for repair of muscle membranes after damage. Mutations in dysferlin lead to a progressive muscular dystrophy. Recent studies suggest additional roles for dysferlin. We set out to study dysferlin's protein-protein interactions to obtain comprehensive knowledge of dysferlin functionalities in a myogenic context. We developed a robust and reproducible method to isolate dysferlin protein complexes from cells and tissue. We analyzed the composition of these complexes in cultured myoblasts, myotubes and skeletal muscle tissue by mass spectrometry and subsequently inferred potential protein functions through bioinformatics analyses. Our data confirm previously reported interactions and support a function for dysferlin as a vesicle trafficking protein. In addition novel potential functionalities were uncovered, including phagocytosis and focal adhesion. Our data reveal that the dysferlin protein complex has a dynamic composition as a function of myogenic differentiation. We provide additional experimental evidence and show dysferlin localization to, and interaction with the focal adhesion protein vinculin at the sarcolemma. Finally, our studies reveal evidence for cross-talk between dysferlin and its protein family member myoferlin. Together our analyses show that dysferlin is not only a membrane repair protein but also important for muscle membrane maintenance and integrity.
authors.affiliation: Center for Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Antoine
authors.initials: A
authors.lastname: de Morrée
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hensbergen
authors.forename: Herman H H B M
authors.initials: HH
authors.lastname: van Haagen
authors.forename: Irina
authors.initials: I
authors.lastname: Dragan
authors.forename: André M
authors.initials: AM
authors.lastname: Deelder
authors.forename: Peter A C
authors.initials: PA
authors.lastname: 't Hoen
authors.forename: Rune R
authors.initials: RR
authors.lastname: Frants
authors.forename: Silvère M
authors.initials: SM
authors.lastname: van der Maarel
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D016000:Cluster Analysis; D000073939:Dysferlin; D020869:Gene Expression Profiling; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012508:Sarcolemma / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: C497505:Dysf protein, mouse; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins; D020543:Proteome; C521810:myoferlin protein, mouse