A 3-D reconstruction of smooth muscle alpha-actinin by CryoEm reveals two different conformations at the actin-binding region.
Cryoelectron microscopy was used to obtain a 3-D image at 2.0 nm resolution of 2-D arrays of smooth muscle alpha-actinin. The reconstruction reveals a well-resolved long central domain with 90 degrees of left-handed twist and near 2-fold symmetry. However, the molecular ends which contain the actin binding and calmodulin-like domains, have different structures oriented approximately 90 degrees to each other. Atomic structures for the alpha-actinin domains were built by homology modeling and assembled into an atomic model. Model building suggests that in the 2-D arrays, the two calponin homology domains that comprise the actin-binding domain have a closed conformation at one end and an open conformation at the other end due to domain swapping. The open and closed conformations of the actin-binding domain suggests flexibility that may underlie Ca2+ regulation. The approximately 90 degrees orientation difference at the molecular ends may underlie alpha-actinin's ability to crosslink actin filaments in nearly any orientation.
Liu
Jun
J
Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380, USA.
Taylor
Dianne W
DW
Taylor
Kenneth A
KA
D000185:Actinin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002147:Calmodulin / Q000737:chemistry; D002645:Chickens; D020285:Cryoelectron Microscopy; D016189:Dystrophin / Q000737:chemistry; D009130:Muscle, Smooth / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation
D000199:Actins; D002147:Calmodulin; D016189:Dystrophin; D000185:Actinin; D002118:Calcium