Discussion_ribosome data

Hi
I have attached an image of ribosome 3D model. This model is a complex of ribosome and 199-base long RNA. In the image you will see string of RNA is coming from the ribosome (if I am not overestimating) but when I look at the refined model, I don’t see any extra density for this RNA.

Is there any way to extract, refine particles with a bigger mask in order to see the extra density? OR any other way that you could kindly suggest to do focused classification , that will be highly appreciated.

Kind Regards
Saif70S_ribosome

Hi @Saif - which bit on this picture do you think is RNA? just to get an idea of what you are trying to focus on

Oli

Hi Oli
I have circled the region by red line.

Cheers
Saif

when you say you don’t see it in the refined map, have you looked at the map filtered to low resolution? say 10-12Å? the region that you have circled looks very mobile

You are right, binding of RNA seems to be very transient. I have attached the image of refined map from two different angles. I only see little bit of RNA density allowing me to model only 10-12 bases. Also tRNA at P and A site are missing. When I did focused classification in relion I can see all 3 tRNAs at E, P, and A sites.
Could you please give me any suggestion how I can see more of it?

Cheers