Hi Julia,

Certainly it is possible that those are false IDs, but it depends as
it often does on other factors.

Did you find other confidently IDed peptides that are not
Phosphorylated?  What is the relative sizes of the positive and
negative mixture distributions in the data?  Do you expect that most
of your correct peptides are phosphorylated?  You can try to enable
the PeptideProphet option "Leave alone all entries with asterisked
score values", which is often helpful in these situations where you
would expect many homologous top matches due to phosph-site
uncertainty.  However, if that doesn't help your situation, the there
maybe other potential points of failure to identify phospho peptides
in this approach.

If you need further help, please compress your data (and database)
into one zip file and then post it somewhere we can download and look
at it (e.g. 
http://tools.proteomecenter.org/wiki/index.php?title=TPP:Frequently_Asked_Questions#How_do_I_upload_files_to_the_SPC_tools_team.3F)

Thank you,

-David

On Fri, Oct 21, 2011 at 12:08 PM, [email protected]
<[email protected]> wrote:
> Hey Guys,
>
> I have a question regarding identification of phosphopeptide using
> TPP. I got a data using an LTQ-XL (using CID) and I ‘ve searched using
> MASCOT search engine (phosphorylation as a variable modification in S/
> T/Y) .  I found more than 50 phosphorylated proteins however when I
> use TPP to look at these phosphorylations I found none! I am wondering
> if all the phoshphorylation detected by MASCOT are in fact false
> positives.
> Here is what I’ve done:
> I use TPP v4.4 rev 1 in W7
> Run MASCOT
> Convert *.dat to *.pepxml
> Run PeptideProphet (I’ve selected the icon “Use phospho information”)
> and ProteinProphet afterwards.
> Can anyone help me?
> Thanks in advance!
>
> Julia
>
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