Fig. 2: Linker remodelling in Rea1ΔAAA2H2α.

The analysis of the ATP, ADP, AMPPNP and ATPγS data sets indicates that linker remodelling in Rea1ΔAAA2H2α is highly similar to linker remodelling in Rea1wt. Like in the case of Rea1wt, ATP-hydrolysis is required to engage the linker with the AAA+ ring. The latter point is also supported by the Rea1ΔAAA2H2α Walker-B mutant, which is impaired in ATP-hydrolysis activity and does not show the AAA+ ring engaged 2D class averages. Unlike Rea1wt, Rea1ΔAAA2H2α is able to sample state 8 in the presence of the slowly hydrolysable ATP analogue ATPγS. White arrow heads highlight a connection between the linker top and the AAA+ ring. Percentage numbers indicate how many particles of the corresponding data set sorted into the displayed 2D class averages.