If you applied a fixed modification during the search then "all" amino
acids of that sort will be modified.  Not all search engines might report
the modification info correctly.  Can you share an example pepXML from your
output of different search engines?

Thank you,

-David


On Fri, Apr 18, 2014 at 11:30 AM, Ruby <[email protected]> wrote:

> Hi-
> I'm processing data with both Sequest and X!Tandem followed by peptide &
> protein prophet. I've used TPP v4.6.3 and v7 and am seeing same issue for
> both versions and for both search engines. I did include a
> reduction/alkylation step during sample prep and I am using C57 as a fixed
> modification during the search, but yet in the results, most of the
> peptides that contain a C are not modified. I'm confused as I thought
> 'fixed' meant it would be required to be present in order for the peptide
> to be identified.  When searching the same data with Morpheus, it is in
> general finding a lot more peptides with the carbamidomethlyation for the
> same protein than I"m getting in my current processing for Sequest &
> Tandem.
>
> This is how my X!Tandem params file is setup:
>
> <note>residue modification parameters</note> <note type="input"
> label="residue, modification mass">57.021464@C</note>
>
> Can anyone clarify whether 'fixed' really means that the modification is
> required to be present if that amino acid is present in the sequence?  Any
> suggestions on where to start looking for my error in the setup?
>
> Thanks!
> Rebekah
>
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