Hi Jesse,

I am not certain I fully understand the issue, but it sounds to me like you
have multiple searches and should therefore have multiple pepXML files
(from you heavy and light).  I would suggest you modify your pipeline
slightly by analyzing the heavy and the light searches separately and
combining them only after PTMProphet has processed the results.  Is this
feasible?

Thanks,
-David

On Mon, Dec 15, 2014 at 2:28 PM, Jesse <[email protected]> wrote:
>
> David,
>
> I think I need to have some modifications added to the TPP code for
> PTMprophet to work.  I trying to combine two database searches that specify
> either fixed light lysine or fixed heavy lysine (+8.0142), both with
> variable modification of diGlycine (+114.0429).  Therefore, lysine in my
> datasets is found modified with +8.0142, +114.0429, or +122.0572.  If you
> add these to TPP, will I be able to combine these searches and validate the
> mods with PTMprophet?
>
> Any help would be appreciated.
>
> Thank you,
> 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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>>>>>> an
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>>>>>> To post to this group, send email to [email protected].
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>>>>>> For more options, visit https://groups.google.com/d/optout.
>>>>>>
>>>>> --
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>>
>
> --
> Jesse Meyer
> Graduate Student
> Department of Chemistry and Biochemistry
> University of California, San Diego
> La Jolla, CA 92092
>
>
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