AAA+ Unfoldase

Dataset

This AAA+ Unfoldase dataset was kindly provided to us by John Rubinstein . The original paper on this dataset was published in Elife 2017, entitled Structure of a AAA+ unfoldase in the process of unfolding substrate .

Learning objectives

2D classification & multi-class ab-initio reconstruction.

Title Value
Title
Description Leave empty
Directory
Create Workspace Enable

Parameter Value
Particle meta path

This should take less than 10 seconds.

Parameter Value
Circular mask diameter (A)
Number of GPUs to parallelize

Choose a set of 2D classes that look sharp and display secondary structure features. Make sure not to exclude classes showing the active state of the unfoldase, as we will use these downstream in ab-initio! However, you should exclude classes that converged to two copies of the "side view", as these particles are from crowded regions of the micrograph.

You should end up with around 133K particles.

Parameter Value
Number of Ab-Initio classes

Inputs Source
Particle stacks Particles from the more populated, inactive, 6-fold symmetric state
Initial volume Use the corresponding volume class
Parameter Value
Symmetry

Launch this job in parallel with the previous homogeneous refinement (using particles from ab-initio reconstruction).

Inputs Source
Particle stacks Particles from the less populated, active, asymmetric state
Initial volume Use the corresponding volume class

Use default parameters