I have tons of high quality krios data collected on a two-component computationally designed de novo protein that has substantial movement engineered into it. In 2D averages both components are visible, however in 3D only one component is ever able to be resolved to side chain detail.
I attended Ali’s webinar on 3DVA and have read the tutorials. The output only ever shows blobs of density for the unresolvable component. It’s frustrating because it’s visible in 2D and so I suspect it must be some processing issue. I have tried every conceivable trick including subtracting signal for the resolved component but still cannot resolve the other component. I have also tried the various output options in the 3DVA display to no avail.
I have several designs where I’m running into this problem. I wonder if it is a good test case for 3DVA improvement? This is an extremely challenging problem and would love to hear any suggestions and/or if there is any time or interest to collaborate on such problems.