Ok, thanks! Just trying to figure out best practices.
Best,
Janel
ᐧ

On Tue, Aug 13, 2019 at 2:45 PM David Shteynberg <
[email protected]> wrote:

> Yes, it makes sense to always run iProphet, to combine the results at the
> peptide level before your combined them at the protein level with
> ProteinProphet.  Your best bet is to combine your PeptideProphet analyses
> into a single analysis with iProphet and then run ProteinProphet on one
> iProphet output.  This may not always be possible given available computer
> resources.  In this case, you can break your data up into several iProphet
> analyses and combine these into one ProteinProphet analysis.  It doesn't
> matter much when you run PTMProphet, but if you run it after iProphet it
> might run a bit faster since it will only compute the localization of the
> combined results.
>
> Cheers,
> -David
>
> On Sat, Aug 10, 2019 at 11:28 AM Janel Beckley <[email protected]>
> wrote:
>
>> Hi David,
>> I just needed to have the exact mass for the K mod. Now it works without
>> errors. Thanks!
>>
>> I have a couple more questions...about file merging in the TPP. If I want
>> to combine a series of MS experiments each with multiple files, I presume I
>> should always run iprophet to combine files prior to running
>> proteinprophet, right? Also, does it matter if PTMprophet is run before or
>> after iprophet? Thanks for all your help!
>>
>> Best,
>> Janel
>> ᐧ
>>
>> On Fri, Aug 9, 2019 at 7:41 PM David Shteynberg <
>> [email protected]> wrote:
>>
>>> Hello Janel,
>>>
>>> Thank you for trying out PTMProphet.   An internal check in the code
>>> verifies that the mod mass during the search matches the mod mass at the
>>> time of running PTMProphet.   Can you verify the 114 on K is in fact the
>>> mass you used at search time?  Can you look at that specific spectrum in
>>> the pepxml file and all check modification listed for that search_hit?
>>>
>>> Thanks,
>>> David
>>>
>>> On Fri, Aug 9, 2019, 4:14 PM Janel <[email protected]> wrote:
>>>
>>>> Hi,
>>>>
>>>> First off I'll just say that it's my first time using PTM Prophet and
>>>> I'm excited to see what it can do! I'm running
>>>> Comet->peptideprophet->ptmprophet in TPP v 5.2.0 and unfortunately, I'm
>>>> running into a number of these errors:
>>>>
>>>> (example)
>>>> ERROR: Cannot initialize for sequence: nKVIDGIDPPK[242], unknown mods
>>>> may exist in spectrum Rep1_wt_010.12231.12231.1
>>>> Please specify modification and rerun PTMProphet ...
>>>>
>>>>
>>>> Here's the command I used for PTMProphet:
>>>> *EXECUTING: cd c:/Users/.../prophet-testing-HBH/Test && c: &&
>>>> C:/TPP/bin/PTMProphetParser STY:79.966,MW:15.995,K:114,n:42.010565
>>>> MINPROB=0.5 MAXTHREADS=0 FRAGPPMTOL=500
>>>> c:/Users/.../prophet-testing-HBH/Test/interact.ipro.pep.xml
>>>> interact.ptm.ipro.pep.xml *
>>>>
>>>> It looks like PTMProphet isn't reading the mass of the modification
>>>> correctly, but it isn't all the time (i.e. some of the modifications are
>>>> analyzed even though they are also the total mass of the aa instead of just
>>>> the mass change). I also tried using high and low mass accuracy files
>>>> and changing the FRAGPPMTOL, but still got the same errors. I must be
>>>> missing something, but I'm not sure what. An option? Something else? Any
>>>> help would be appreciated!
>>>>
>>>> Thanks in advance,
>>>> Janel
>>>>
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