Sorry I got cut off.  You can find the full parameter file I used
here:  https://dl.dropboxusercontent.com/u/21286225/tandem_params.xml


Next I had to modify PTMProphetParser.exe to be able to process this
type of data.  The updated binary can be found here:
https://dl.dropboxusercontent.com/u/21286225/PTMProphetParser.exe

After rerunning the search and xinteract with PeptideProphet and
iProphet and verifying that I was getting good matches, I used the
following settings for PTMProphetParsers:

 MZTOL=0.4 STY,79.966,M,15.995,nK,6.0318

I hope this helps you process your data on your own.  If you still
cannot get it to work based on this I can send you the actual pepXMl
results as well.

Thanks for continuing to use the TPP and for helping us improve the tools!

-David

On Tue, Aug 26, 2014 at 4:35 PM, David Shteynberg
<[email protected]> wrote:
> Dear Oded,
>
> Sorry it has taken me this long to work the bugs out in this analysis.
> The first problem I found was with your search parameters.   Instead
> of using two different static mods on K and n-terminus (which is not
> supported by TPP in a single search),  I used a static mod in
> combination with a variable mod so instead of:
>          <parameter name="residue, modification mass"
> value="+57.021464@C,+28.031300@K,+28.031300@["/>
>          <parameter name="residue, modification mass 1"
> value="+57.021464@C,+34.063117@K,+34.063117@["/>
>          <parameter name="residue, potential modification mass"
> value="15.994915@M,79.966331@Y,79.966331@S,79.966331@T"/>
>
> we now have:
>
>
>
>
> On Tue, Aug 12, 2014 at 1:19 PM, Jesse Meyer <[email protected]> wrote:
>> Even though the cysteine mod is variable, I was able to successfully run
>> PTMprophet by entering the cysteine mod as one of the PTMs for PTMprophet.
>> Do you think this will cause problems?  Also, what localization score should
>> be used as a cutoff for correct localization?
>>
>> Thanks,
>> Jesse
>>
>>
>> On 8/12/2014 1:16 PM, David Shteynberg wrote:
>>>
>>> You need to only enter the variable mods for PTMProphet.  However,
>>> mods that are unknown will not be recognized without a change to the
>>> code.  A future version of PTMProphet will hopefully correct these
>>> issues.
>>>
>>> -David
>>>
>>> On Tue, Aug 12, 2014 at 11:59 AM, Jesse Meyer <[email protected]>
>>> wrote:
>>>>
>>>> I am also getting this error with a peptide containing a fixed
>>>> modification
>>>> of cysteine (N-ethylmaleimide +125.047679).  I am running PTMprophet to
>>>> localize a different variable modification on lysine.   Do I need to
>>>> enter
>>>> all mods including fixed mods under PTMprophet in the Petunia GUI?
>>>>
>>>> Thanks,
>>>> Jesse
>>>>
>>>>
>>>> On Saturday, July 12, 2014 7:13:57 AM UTC-7, [email protected] wrote:
>>>>>
>>>>> Sure. Let me know what you need and I will send you the data.
>>>>> Best,
>>>>> Oded
>>>>>
>>>>> On Saturday, July 12, 2014 5:11:39 AM UTC+10, David Shteynberg wrote:
>>>>>>
>>>>>> Hi Oded,
>>>>>>
>>>>>>
>>>>>> It is likely there is an issue with PTMProphet that doesn't recognize
>>>>>> the modifications in your dataset.  If you can share a portion of your
>>>>>> data for testing purposes I can help troubleshoot further.
>>>>>>
>>>>>> Thank you!
>>>>>> -David
>>>>>>
>>>>>> On Tue, Jul 1, 2014 at 5:33 AM, Oded <[email protected]> wrote:
>>>>>>>
>>>>>>> Hi all,
>>>>>>> I am using TPP 4.7.1 on Win7 and currently trying to analyze data of
>>>>>>> dimethylated phosphopeptides searched with X!tandem.
>>>>>>> The searches were done by using 2 sets of fixed modifications for the
>>>>>>> Cys
>>>>>>> alkylation plus light or heavy dimethylation on N-term and K  with STY
>>>>>>> phosphorylation, Met-Ox and protein N-term acetylation as set as
>>>>>>> variable
>>>>>>> modifications.
>>>>>>> When I try to run PTM prophet (following peptide prophet and iprophet
>>>>>>> runs)
>>>>>>> I getting the following error:
>>>>>>>
>>>>>>>
>>>>>>> EXECUTING: run_in
>>>>>>> c:/Inetpub/wwwroot/ISB/data/Oded/Enzo/Phospho_June2014;
>>>>>>> c:\Inetpub\tpp-bin\PTMProphetParser
>>>>>>> STY,79.966,C,57.021464,n,34.063117,K,34.063117,M,15.99 MZTOL=0.1
>>>>>>>
>>>>>>>
>>>>>>> c:/Inetpub/wwwroot/ISB/data/Oded/Enzo/Phospho_June2014/interactNative1.ipro.pep.xml
>>>>>>> interactNative
>>>>>>> INFO: Writing file interactNative1.ptm.pep.xml ...
>>>>>>> INFO: Reading file
>>>>>>>
>>>>>>>
>>>>>>> c:/Inetpub/wwwroot/ISB/data/Oded/Enzo/Phospho_June2014/interactNative1.ipro.pep.xml
>>>>>>> ...
>>>>>>> WARNING: Cannot initialize for sequence: n[71]M[147]KDEPRSTNLFMK,
>>>>>>> unknown
>>>>>>> mods may exist in spectrum CHH20140618_M1HTiO2_100mM.03738.03738.2
>>>>>>> WARNING: Cannot initialize for sequence: n[35]K[162]S[167]PAAAR,
>>>>>>> unknown
>>>>>>> mods may exist in spectrum CHH20140618_M1HTiO2_100mM.03900.03900.2
>>>>>>> WARNING: Cannot initialize for sequence: n[35]VPS[167]RHINIGR, unknown
>>>>>>> mods
>>>>>>> may exist in spectrum CHH20140618_M1HTiO2_100mM.04358.04358.2
>>>>>>> WARNING: Cannot initialize for sequence:
>>>>>>> n[35]EGEEPT[181]VYSDEEEPK[162]DESAR, unknown mods may exist in
>>>>>>> spectrum
>>>>>>> CHH20140618_M1HTiO2_100mM.04499.04499.3
>>>>>>> WARNING: Cannot initialize for sequence: n[35]GTPGPAVR, unknown mods
>>>>>>> may
>>>>>>> exist in spectrum CHH20140618_M1HTiO2_100mM.05584.05584.2
>>>>>>>      WARNING: Unrecognized mod on peptide: n[35]SCFESS[167]PDPELK[162]
>>>>>>>      WARNING: Unrecognized mod on peptide: n[35]SCFESS[167]PDPELK[162]
>>>>>>>      WARNING: Unrecognized mod on peptide: n[35]SCFESS[167]PDPELK[162]
>>>>>>>      WARNING: Unrecognized mod on peptide: n[35]SCFESS[167]PDPELK[162]
>>>>>>>      WARNING: Unrecognized mod on peptide: n[35]SCFESS[167]PDPELK[162]
>>>>>>>      WARNING: Unrecognized mod on peptide: n[35]SCFESS[167]PDPELK[162]
>>>>>>>      {this appear 100-200 times}
>>>>>>>
>>>>>>>
>>>>>>>
>>>>>>> This application has requested the Runtime to terminate it in an
>>>>>>> unusual
>>>>>>> way.
>>>>>>> Please contact the application's support team for more information.
>>>>>>>
>>>>>>> command "c:\Inetpub\tpp-bin\PTMProphetParser
>>>>>>> STY,79.966,C,57.021464,n,34.063117,K,34.063117,M,15.99 MZTOL=0.1
>>>>>>>
>>>>>>>
>>>>>>> c:/Inetpub/wwwroot/ISB/data/Oded/Enzo/Phospho_June2014/interactNative1.ipro.pep.xml
>>>>>>> interactNative1.ptm.pep.xml" failed: No such process
>>>>>>>
>>>>>>>
>>>>>>> I tried adding both heavy and light dimethylation (+28 and +34 on K
>>>>>>> and
>>>>>>> N-term) just light and just heavy (as shown above), the the mass of
>>>>>>> the
>>>>>>> DiMet N-term as it appear in the pepXML (+35 or +29)  but the error is
>>>>>>> still
>>>>>>> the same...
>>>>>>> Is there a way to sort this?
>>>>>>> Many thanks,
>>>>>>> Oded
>>>>>>>
>>>>>>> --
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>>>>
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>>
>>
>>
>> --
>> Jesse G. Meyer
>> Ph. D. Candidate
>> Department of Chemistry and Biochemistry
>> University of California, San Diego
>> [email protected]
>>
>>
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