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Mol Biol Cell ; 25(19): 3037-48, 2014 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-25143396

RESUMEN

Dysferlin and calpain are important mediators of the emergency response to repair plasma membrane injury. Our previous research revealed that membrane injury induces cleavage of dysferlin to release a synaptotagmin-like C-terminal module we termed mini-dysferlinC72. Here we show that injury-activated cleavage of dysferlin is mediated by the ubiquitous calpains via a cleavage motif encoded by alternately spliced exon 40a. An exon 40a-specific antibody recognizing cleaved mini-dysferlinC72 intensely labels the circumference of injury sites, supporting a key role for dysferlinExon40a isoforms in membrane repair and consistent with our evidence suggesting that the calpain-cleaved C-terminal module is the form specifically recruited to injury sites. Calpain cleavage of dysferlin is a ubiquitous response to membrane injury in multiple cell lineages and occurs independently of the membrane repair protein MG53. Our study links calpain and dysferlin in the calcium-activated vesicle fusion of membrane repair, placing calpains as upstream mediators of a membrane repair cascade that elicits cleaved dysferlin as an effector. Of importance, we reveal that myoferlin and otoferlin are also cleaved enzymatically to release similar C-terminal modules, bearing two C2 domains and a transmembrane domain. Evolutionary preservation of this feature highlights its functional importance and suggests that this highly conserved C-terminal region of ferlins represents a functionally specialized vesicle fusion module.


Asunto(s)
Calpaína/metabolismo , Membrana Celular/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas Musculares/metabolismo , Sinaptotagminas/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas de Unión al Calcio/metabolismo , Calpaína/genética , Proteínas Portadoras , Células Cultivadas , Disferlina , Células HEK293 , Células Endoteliales de la Vena Umbilical Humana , Humanos , Fusión de Membrana/fisiología , Proteínas de la Membrana/genética , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Proteínas Musculares/genética , Isoformas de Proteínas , Estructura Terciaria de Proteína , Alineación de Secuencia , Proteínas de Motivos Tripartitos
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