Actin interaction with purified dystrophin from electric organ of Torpedo marmorata: possible resemblance with filamin-actin interface.
We have purified dystrophin from Torpedo marmorata electric tissue by means of alkaline extraction in conjunction with an affinity chromatography column using anti-peptide antibodies. Using solution (cosedimentation) and solid phase experiments (sedimentation with Sepharose filamentous actin and ELISA), we have demonstrated that purified dystrophin is able to bind filamentous and monomeric actin. Using ELISA coupled with biotin labelled peptides and taking advantage of strong affinity binding of streptavidin-biotin complex, we have identified two exposed sequences of the actin molecule implicated in dystrophin binding: fragment 40-113, further restricted to peptide 75-106 and peptide 360-372. In a previous study, we have shown that fragment 40-113 displays binding site(s) for filamin but probably not for alpha-actinin. Moreover, we have recently reported that alpha-actinin and filamin display divergent behaviours towards conformational changes of actin. In this study, we have demonstrated that, similarly to filamin, dystrophin binding is insensitive to the locking of actin in a monomeric conformation. Taken together, these results lead us to favour the idea that dystrophin could share properties in common with filamin in its binding of actin.
Lebart
M C
MC
Centre de Recherche de Biochimie Macromoléculaire, U. 249 Institut National de la Santé et de la Recherche Médicale, Laboratoire de Recherche sur la Motilité Cellulaire, Ecole Pratique des Hautes Etudes, Université de Montpellier I, BP 5051, France.
Casanova
D
D
Benyamin
Y
Y
D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002352:Carrier Proteins / Q000378:metabolism; D003285:Contractile Proteins / Q000378:metabolism*; D016189:Dystrophin / Q000302:isolation & purification / Q000378:metabolism*; D004557:Electric Organ / Q000502:physiology*; D064448:Filamins; D008840:Microfilament Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D014101:Torpedo / Q000378:metabolism*
D000199:Actins; D002352:Carrier Proteins; D003285:Contractile Proteins; D016189:Dystrophin; D064448:Filamins; D008840:Microfilament Proteins; D010455:Peptides; D000185:Actinin