His-tag peptides

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His-tag peptides jung165  2020-06-04 21:12
 

Hi Skyline team.
I always appreciate for the fascinating tool that is applicable for targeted protemics!

I have one quick question about selecting peptides.
I prepared the his-tag purified and enriched proteins which are transfected in Ecoli.
Therefore, I would like to insert modification about His-tag on my target peptide but, in modification tap I could not find such modification.
Is it applicable to just put 6H(histidine) after my target peptide and inset on Skyline??

I really appreciate for your help
Sincerely,

 
 
Mike MacCoss responded:  2020-06-04 23:02

Thanks for the post. When we have studied fusion proteins we add an altered protein sequence, not a modification to the peptide. For example if you have a peptide sequence

IAMAPEPTIDEK

You could have a 6 His on the N-term

HHHHHHIAMAPEPTIDEK

On the c-term it would be

IAMAPEPTIDEKHHHHHH

In the later case the 6 his would be cleaved off by trypsin but hopefully you get the idea.

Cheers,
Mike

 
jung165 responded:  2020-06-04 23:35

It actually helped me a lot
Really appreciate for that!

Sincerely,