[3dem] This is my question to all the members experts and non-experts who are working on membrane proteins in the CryoEM

[Guillaume Gaullier]

Hi Aninda,

Here is a recent review on cryoEM of small proteins: https://urldefense.com/v3/__https://doi.org/10.1016/j.str.2024.03.003__;!!Mih3wA!HjneFU3_I0LNA5Ecsh4kLh0y-aemd0SeU9_j156zgWRzsjVBc3UjwUKC4a_IbFBNlxbLfRt0INvqDSUWJasoXaTuv7SrZlB_$ 
It is not specific to membrane proteins, but might answer some of your questions or give you pointers to look further.

This other article with a systematic exploration of data collection parameters for GPCRs might be more directly relevant to you, even though GPCRs are larger than the proteins you describe: https://urldefense.com/v3/__https://doi.org/10.1038/s41467-021-24650-3__;!!Mih3wA!HjneFU3_I0LNA5Ecsh4kLh0y-aemd0SeU9_j156zgWRzsjVBc3UjwUKC4a_IbFBNlxbLfRt0INvqDSUWJasoXaTuv0toUZGm$ 

When you don’t get answers on this 3DEM list, I suggest to also search the archives of the CCPEM list: https://urldefense.com/v3/__https://www.jiscmail.ac.uk/cgi-bin/webadmin?A0=CCPEM__;!!Mih3wA!HjneFU3_I0LNA5Ecsh4kLh0y-aemd0SeU9_j156zgWRzsjVBc3UjwUKC4a_IbFBNlxbLfRt0INvqDSUWJasoXaTuv7veKZys$ 
And if you still don’t find anything relevant, of course you are welcome to post your questions to the CCPEM list as well. Some people follow both lists (I do), but I don’t know how much the two populations overlap, so for certain questions you might reach more people with potential answers on the other list.

I hope this helps,

Guillaume


On 6 Apr 2024, at 22:46, Aninda Dutta <anindadutta86 at gmail.com<mailto:anindadutta86 at gmail.com>> wrote:

Dear 3DEM list members,
I have two small bacterial membrane enzymes (sub-50 kDa) that I want to work with. Both of them are known to dimerize under certain conditions and I want to understand the conformational dynamics of these systems under native lipid environments (Liposome, nanodiscs) using CryoEM.
Regarding that, I have some questions to ask all of you -

  1.
How are we as a community doing with sub-50 kDa CryoEM without using Fabs or other nanobodies?
  2.
What are the challenges, and can we come over it?
  3.
Tell me any suggestions or tips you may have before starting this project.

Last but not the least, I am open to any interesting collaborations.

Best,
Aninda

---------------------
Aninda Dutta
UVA Chem Graduate Student
University of Virginia
Charlottesville, VA

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