Dystrophin and utrophin have distinct effects on the structural dynamics of actin.
We have used time-resolved spectroscopy to investigate the structural dynamics of actin interaction with dystrophin and utrophin in relationship to the pathology of muscular dystrophy. Dystrophin and utrophin bind actin in vitro with similar affinities, but the molecular contacts of these two proteins with actin are different. It has been hypothesized that the presence of two low-affinity actin-binding sites in dystrophin allows more elastic response of the actin-dystrophin-sarcolemma linkage to muscle stretches, compared with utrophin, which binds via one contiguous actin-binding domain. We have directly tested this hypothesis by determining the effects of dystrophin and utrophin on the microsecond rotational dynamics of a phosphorescent dye attached to C374 on actin, as detected by transient phosphorescence anisotropy (TPA). Binding of dystrophin or utrophin to actin resulted in significant changes in the TPA decay, increasing the final anisotropy (restricting the rotational amplitude) and decreasing the rotational correlation times (increasing the rotational rates and the torsional flexibility). This paradoxical combination of effects on actin dynamics (decreased amplitude but increased rate) has not been observed for other actin-binding proteins. Thus, when dystrophin or utrophin binds, actin becomes less like cast iron (strong but brittle) and more like steel (stronger and more resilient). At low levels of saturation, the binding of dystrophin and utrophin has similar effects, but at higher levels, utrophin caused much greater restrictions in amplitude and increases in rate. The effects of dystrophin and utrophin on actin dynamics provide molecular insight into the pathology of muscular dystrophy.
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
Ewa
E
Prochniewicz
Davin
D
Henderson
James M
JM
Ervasti
David D
DD
Thomas
D000199:Actins / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D016189:Dystrophin / Q000502:physiology*; D007313:Insecta; D007700:Kinetics; D008954:Models, Biological; D008956:Models, Chemical; D008958:Models, Molecular; D009136:Muscular Dystrophies / Q000378:metabolism; D010758:Phosphorus / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D049411:Utrophin / Q000737:chemistry / Q000502:physiology*
D000199:Actins; D016189:Dystrophin; D049411:Utrophin; D010758:Phosphorus