Hello,
I have a small purified protein, 15kDa. Iam seeking some technical service to build its 3D structure by Cryo -Sem, or NMR. However, I was told by some technicians that Cryo-Sem is only suitable for big protein with MW>50kDa. Very confused.
No modeling in silicon, because I couldn't find a suitable template in the database.
Can NWR be used for an isolated native protein structure building?
Any suggestions?
Zha
Hi,
For protein structure determination by NMR a high concentration of your purified target protein will be needed. Also, your protein need to be labeled with deuterium, N15 and C13. Please, check this publication doi.org/10.1021/bi0118308. For Cryo-EM of small proteins people are trying to get a purified complex of target protein with specific antibodies ou liked to other proteins like MBP commonly used for recombinant expression in E. coli. You can try to express your target protein in E coli, purify a high concentration of it and go to crystallization. Thus, NMR or crystallization will be more suitable for your target protein of 15 kDa. Good luck.
I think there is a commercial service out there like creative biostructure. NMR structure determination is not an easy task in general and it is expensive. If you have enough manpower in your group and also NMR time, you can try: https://www.youtube.com/playlist?list=PLmYqoarOFAZ-1z6LRDpgUMVqKZOxqohyT
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