TY  - GEN
N2  - Subject of these studies was the strong binding complex of actin and myosin V. How the molecular
motor protein myosin powers muscles by generating mechanical force out of chemical energy at high
ef?ciency fascinates researchers already for a long time. Myosin V offers the unique possibility to
study not only the rigor state but also its preceding strong binding ADP state, which is only transiently
present in other myosins. Since the actomyosin complex could not be studied by protein crystallography so far, transmission electron microscopy (TEM) of vitri?ed protein complexes is the method
of choice. New reconstruction strategies were applied, to enhance the interpretable resolution in 3D
electron microscopy, together with subsequent molecular dynamics (MD) simulations of protein crystal structures. Such a combined approach allows for almost the same resolution as X-ray diffraction
facilitates. As a new ? and in principle superior ? reconstruction scheme a ?ltered least squares re-
construction algorithm was applied in order to obtain a better de?ned density localization. Thorough
analysis combined with electron tomography studies revealed ?exible specimen properties that limit
the applicability of the current implementation of the least squares method due to necessary ad hoc
assumptions. As a ?rst step to reduce these assumptions iterative helical reconstruction was applied
and yielded densities with reliable 8 Å resolution. Following MD simulations were able to resolve
functional differences of myosin between the rigor and the strong binding ADP state.

TI  - Application of new reconstruction strategies to enhance the interpretable resolution in 3D Electron Microscopy - Studies of Rigor and ADP binding complexes of Actin and Myosin V

UR  - https://archiv.ub.uni-heidelberg.de/volltextserver/16828/
AV  - public
ID  - heidok16828
A1  - Wulf, Sarah Elise Freyja
Y1  - 2014///
ER  -