Hello Piero,

It appears that the version of PTMProphet in Philosopher is a bit out of
date.  The latest TPP version is 5.1.0, and PTMProphet has been updated
with bugfixes and new features in the meantime. I have run my latest
PTMProphet code (targeted for the 5.2.0 version of TPP) on your data.  For
the PSMs matching this peptide I cannot replicate the issue you've
reported.  Instead, PTMProphet seems to be positioning the mass
modification on the Asparagine in your peptide as you should see in the
following result file:

https://www.dropbox.com/s/z3xf379ulljr3rr/01928_GA1_P020123_S00_R01_R1.PTMPro.pep.xml?dl=0

Thanks for your interest in PTMProphet.

Cheers!

-David

On Fri, May 18, 2018 at 8:40 AM, Piero Giansanti <[email protected]>
wrote:

> Hello David,
>
> I am using PTMprophet via philosopher, and from the xml output it seems it
> is this version
> TPP v5.0.1 Post-Typhoon dev, Build 201706282113-exported (Linux-x86_64)
>
> Regarding the settings, I am using --massdiffmode --mztol 0.01. The mass
> tolerance is not exactly the one used for the database search (20 ppm), but
> it should be somehow close to it.
>
> Here https://ufile.io/wiqqb the mzML, pepXML and the output I generated.
>
> Thanks
>
>
> On Friday, May 18, 2018 at 4:21:27 PM UTC+2, David Shteynberg wrote:
>>
>> Hello Piero,
>>
>> Thank you for trying the software.  This is indeed an unusual result.
>> Can you tell me which version of PTMProphet you are using and how you are
>> running the tool?  Is it possible for you to post your pepXML file and data
>> file online for me to download and test this particular ID?
>>
>> Cheers,
>> -David
>>
>> On Fri, May 18, 2018 at 1:53 AM, Piero Giansanti <[email protected]>
>> wrote:
>>
>>> Hi,
>>>
>>> I am using PTMprophet to localize the set of modifications (i.e. mass
>>> shifts) identified by running an open search with MSfragger, but I have
>>> some doubt regarding the interpretation of the output.
>>> In particular, MSfragger identified this peptide
>>> *GG(ox)MNGASPPTSQK*
>>> with a mass shift of 0.9869 Da, which match to the following
>>> modification according to Unimod
>>> 0.9840:Deamidated (Deamidation), 0.9840:Asn->Asp (Asn->Asp
>>> substitution), 0.9840:Gln->Glu (Gln->Glu substitution).
>>>
>>> Of these 3 only the first 2 are possible (there is no Q in the sequence,
>>> so there cannot be any Gln->Glu substitution).
>>>
>>> Now, the output of PTMprophet is as follows:
>>>
>>> *G(0.000)G(0.000)M(0.000)N(0.000)G(0.000)A(0.000)S(0.000)P(0.000)P(0.000)T(0.000)S(0.000)Q(0.000)K(0.000)*
>>>
>>> Is a probability always 0 somehow an expected behavior? How do I
>>> interpret this? Am I right to say that the 0.9869 Da shift cannot be
>>> localized to any residues?
>>> My concerns comes from the fact that even if the mod cannot be
>>> localized, I would have expected to get a probability of (~0.008, i.e.
>>> 100/length of the peptides) for each residues...
>>>
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>>
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