I'm interested in studying the per-sulphidation change in my protein of interest using LC MS MS. Can anybody recommend methods which does not involve labeling technique.
Hi,
Can you educate me on what per-sulphidation of a protein entails?
Thanks!
Hi,
The cysteine residues in the protein undergo modification from SH to SSH.This can occur due to hydrogen sulfide signalling.
First thing I'd do, due to the much stronger acidity of the SH bond in persulfides compared to thiols, is look at the protein in the negative ionization mode before and after persulfidation. Ideally the SH protein may not show at all, but the SSH-modified one will.
Hi Julius,
Thank you for the reply. I have to do this experiment in IP pull down fraction and perform tryptic digestions. If we change the ionisation mode then we need to change to buffers also right. And another problem is i do not have a recombinant protein for my protein of interest. Pls clarify....
You may have to adjust your buffers, yes. But if/when you have enough material, you may simply try it in slightly basic conditions (negative mode with ammonia added); or, you can even try it in the positive mode with ammonia added, to see if there will be any ammonium adducts.
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