Structural studies on maturing actin filaments
Journal Article

Published: Wednesday, May 25, 2011
Journal: BioArchitecture, Volume 1, Number 3, Pages 127-133
Link: http://www.es.landesbioscience.com/journals/BioArchitecture/article/16714/

Authors (1 total): M. H. Jensen

Abstract:

We have previously reported that actin undergoes a conformational transition (which we named "maturation") during polymerization, and that the actin-binding protein, caldesmon (CaD), when added at an early phase of polymerization, interferes with this process. The pre-transition filament is characterized by relatively low pyrene-fluorescence intensity when pyrene-labeled actin is used as a reporter of subunit assembly into filaments, whereas the mature filament emits a characteristic enhanced fluorescence. Previously reported co-sedimentation experiments suggest that filament formation is not inhibited by the presence of CaD, despite blocking the transition associated with filament maturation. In this study we visualized structural effects of CaD on the assembly of actin filaments by TIRF and electron microscopy. CaD-free actin forms "rough" filaments with irregular edges and indistinct subunit organization during the initial phase (~20 min under our conditions) of polymerization as reported previously by others, which most likely correspond to the pre-transition state preceding the maturation step. Later during the polymerization process "mature" filaments exhibit a smoother F-actin appearance with easily detectible double helically arranged actin subunits. While the inclusion of the actin-binding domain of CaD during actin filament polymerization does not affect the elongation rate, it is associated with a prolonged pre-transition phase, characterized by a delayed alteration (rough to smooth) of the appearance of filaments, consistent with a later onset of the maturation process.