The proper way to run PTMProphet is with the mass shift and grouping amino
acids sharing that shift.  STY,79.966 is correct.  Are you using any other
mods in your search...variable or static?  A mass mod that is unknown to
PTMProphet is causing the warning, can you look for this peptide ti the
input files to see if it has any other mods on it?

Tolerance should be same as search fragment tolerance.

Do not update will keep the orignal modifications call made by search
algorithm instead of updating to PTMProphet predicted sites.

Prior mean prior probabilty that each site is modified without looking at
peaks but assuming the IDis correct.  If a peptide has 3 sites and one mod
prior is 1/3.

ProteinProphet can be run on PTMProphet containing pepXML file.  The
peptides should display the PTMProphet mod predictions unless the no
upgrade option youasked about is enabled.

Hope this helps,

David
On Dec 11, 2013 11:09 AM, "Vidya Venkatraman" <[email protected]>
wrote:

> Hi David,
>
> I am running the TPP v4.6 OCCUPY rev 3, Build 201307241109 (MinGW)
> version of TPP. I have recently started exploring PTM prophet in TPP. I had
> a few questions regarding the options and parameters within PTM Prophet.
>
> Feel free to point me to any documentation or tutorial if there is one
> already on these parameters. I can also send any input or result files if
> needed.
>
> 1) The default residue mass shift of 79.96 on STY does not seem to work in
> my case. Although, if i give the total mass change of 167 for S, 181 for T
> and 243 for Y, it works. I noticed that my pep.xml files mention total
> masses. Should total mass change be used?
> This is the error I get:
>
> running: "C:/Inetpub/tpp-bin/PTMProphetParser STY,79.966 MZTOL=0.1 
> interact.ipro.pep.xml interact.ptm.ipro.pep.xml"
> INFO: Writing file interact.ptm.ipro.pep.xml ...
> INFO: Reading file interact.ipro.pep.xml ...
>       WARNING: Unrecognized mod on peptide: 
> MAQPAWGLSASHRAPAPPPPPLLLPLPLLLPLLELPLLHRS[167]S
>
> I also noticed that even if i use total mass change, for one peptide, I
> got this warning
>
> running: "C:/Inetpub/tpp-bin/PTMProphetParser S,167,T,181,Y,243 MZTOL=0.1 
> interact.ipro.pep.xml interact.ptm.ipro.pep.xml"
> INFO: Writing file interact.ptm.ipro.pep.xml ...
> INFO: Reading file interact.ipro.pep.xml ...
>       WARNING: Unrecognized mod on peptide: IPESLGLMTLPPESLQKT[181]
> command completed in 1 sec
>
>
> 2) Should the m/z tolerance in the PTM prophet parameters be the same as
> the fragment tolerance used during the search? If not, What does it exactly
> represent?
>
> 3) What is the "Do not update modification_info tags in pepXML" option
> meant for?
>
> 4) Within the interact.ptm.ipro.pep.xml, what does the tag
> "ptmprophet_result prior="0.5" mean?
>
> <analysis_result analysis="ptmprophet">
> <ptmprophet_result prior="0.5" ptm="PTMProphet_T79.9663" 
> ptm_peptide="IPESLGLMT(0.916)LPPESLQKT(0.084)">
> <mod_aminoacid_probability position="9" probability="0.916"/>
> <mod_aminoacid_probability position="18" probability="0.084"/>
> </ptmprophet_result>
>
>
> 5) The current TPP workflow seems to have options to run ProteinProphet
> after PeptideProphet and/or after InterProphet. I was wondering if it was
> possible to run ProteinProphet after PTMProphet? (i.e. can we run
> ProteinProphet on the interact.ptm.ipro.pep.xml file created by
> PTMProphet). Are there any reasons as to why that should not be done or
> cannot be done after PTMProphet?
>
> I would highly appreciate any inputs or suggestions from your end.
>
> Thanks in advance,
> Vidya
>
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