i like this post (click again to cancel)
0
i dont like this post (click again to cancel) remove favorite mark from this question (click again to restore mark)

Dear Friends,

I am not able to figure out how to determine the side chain assignment of Last C-terminal SERINE residue of my protein. I can determine CA, CB, CO, N,H values from HNCA, CBCANH, HNCACO. Can someone tell which experiment will give me the information of HA, HB2 and HB3

Regards

Arun

asked Oct 09 '11 at 06:33

Arun%20Gupta's gravatar image

Arun Gupta
11

updated Oct 09 '11 at 09:16

Evgeny%20Fadeev's gravatar image

Evgeny Fadeev
5571


2 Answers:
i like this answer (click again to cancel)
0
i dont like this answer (click again to cancel)

Hi, You need common side-chain specific experiments like : HC(C)NH, H(C)CH-TOCSY, (H)CCH-TOCSY and HBHANH.

The three first sequences are based on magnetization mixing by spin-lock on carbon frequency and the last one based on polarization transfert from proton to carbone followed by polarization transfert from CA/CB to NH.

H(C)CH-TOCSY and (H)CCH-TOCSY need some frequency to anchor side-chain assignments to backbone assignments. CA, CB, HA (find by common backbone assignment experiments) can be used as anchor-frequency. HC(C)NH and HBHANH correlate directly NH couple to side-chain frequency so assignment is straightforward.

hope that help. YM

link

answered Oct 10 '11 at 03:16

Yoan%20Monneau's gravatar image

Yoan Monneau
136

i like this answer (click again to cancel)
0
i dont like this answer (click again to cancel)

In addition to the experiments suggested by Yoan, a TOCSY-15N-HSQC experiment will correlate the Ha and Hb protons to the backbone amide.

Alternately, if you have all the other Ha, Hb, Ca, and Cb shifts in you protein assigned assigned, that region of a 13C-HMQC (or HSQC) is usually well resolved and you may be able to assign those protons using that spectrum.

link

answered Oct 11 '11 at 09:24

Andrew%20Fowler's gravatar image

Andrew Fowler
96

Your answer
Please start posting your answer anonymously - your answer will be saved within the current session and published after you log in or create a new account. Please try to give a good answer, for discussions, please use comments and please do remember to vote (login to vote)
toggle preview

Tags:

×2
×2

Asked: Oct 09 '11 at 06:33

Seen: 1,350 times

Last updated: Oct 11 '11 at 09:24

powered by CNPROG