The uncoupling of the effects of formins on the local and global dynamics of actin filaments.
In this study, experiments were carried out in the conventional and saturation-transfer electron paramagnetic resonance (EPR) time domains to explore the effect of mDia1-FH2 formin fragments on the dynamic and conformational properties of actin filaments. Conventional EPR measurements showed that addition of formin to actin filaments produced local conformational changes in the vicinity of Cys-374 by increasing the flexibility of the protein matrix in the environment of the label. The results indicated that it was the binding of formin to the barbed end that resulted in these conformational changes. The conventional EPR results obtained with actin labeled on the Lys-61 site showed that the binding of formins could only slightly affect the structure of the subdomain 2 of actin, reflecting the heterogeneity of the formin-induced conformational changes. Saturation transfer EPR measurements revealed that the binding of formins decreased the torsional flexibility of the actin filaments in the microsecond time range. We concluded that changes in the local and the global conformational fluctuations of the actin filaments are associated with the binding of formins to actin. The results on the two EPR time domains showed that the effects of formins on the substantially different types of motions were uncoupled.
Kupi
Tünde
T
Department of Biophysics, Faculty of Medicine, University of Pécs, Pécs, Hungary.
Gróf
Pál
P
Nyitrai
Miklós
M
Belágyi
József
J
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*; D000199:Actins / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology*; D004578:Electron Spin Resonance Spectroscopy; D000080865:Formins; D051379:Mice; D009038:Motion; D009068:Movement / Q000187:drug effects; D012399:Rotation; D013113:Spin Labels; D013696:Temperature; D013997:Time Factors
D000199:Actins; D002352:Carrier Proteins; C491464:Diap1 protein, mouse; D000080865:Formins; D013113:Spin Labels