Hi-
I finally figured out that if I do the mzXML conversion via the TPP prior
to searching (rather than using MSConvert on another system), this seemed
to make a difference in allowing TPP to process data correctly. So that
solution is fixed. However, now I see a new issue.  In older version of
TPP, the NxS/T motif was colored in the ProteinProphet results if I
indicated that prior to proteinprophet processing. Now, I can't get that
button to work. I have it checked in the box at the top of the
ProteinProphet output, but it won't highlight them or calculate the # of
NxS/T peptides as it used to do. (I do a lot of N-glycopeptide capture).

Thanks!
Rebekah

On Mon, Feb 27, 2012 at 12:30 PM, Rebekah Dawson <[email protected]>wrote:

> Hi-
> Thanks for the response. I always have it set to "trypsin" when converting
> the Sequest search results to pepXML within the TPP. Is there another place
> to specify enzyme?  I have the same boxes checked for peptideprophet that I
> always used when going through the pipeline.  One other thing I noticed was
> that in this new version of TPP I have the option of doing a 'semi-enzyme'
> process, which is actually what the original search is using. So I have now
> tried doing the pepXML conversion with straight 'trypsin' or with this
> extra 'semi' box checked also. Both of them create this failure. I have
> also tried it with specifying the sequest.params file during pepXML
> conversion or without specifying it, and get the same error either way. I
> also don't understand where "MALDI mode" comes from in the log listed
> below. I am not using MALDI data and I do not check the "MALDI data" box.
>
> Any help is much appreciated!!
> R
>
>
> On Mon, Feb 27, 2012 at 11:27 AM, [email protected] <
> [email protected]> wrote:
>
>> Please try to rerun the pipeline once specifying the enzyme to see if
>> that helps.
>>
>> Sent from my Android Device
>>
>>
>> ----- Reply message -----
>> From: "Ruby" <[email protected]>
>> To: "spctools-discuss" <[email protected]>
>> Subject: [spctools-discuss] PeptideProphet error in 4.5.2
>> Date: Mon, Feb 27, 2012 7:02 am
>>
>>
>> I have been using TPP 4.0 for over a year with no issues. Last week I
>> upgraded to 4.5.2 and can't get PeptideProphet to work. I"m not doing
>> anything in terms of how I proceed through the pipeline and what
>> settings I use. I get the following error and I get it whether I try
>> to do the pep.XML conversion in this version of TPP, or whether I use
>> pepXML files generated using the previous version of TPP  (and which
>> worked fine in peptideprophet before). Any help would be much
>> appreciated!
>>
>> Command 1 [Mon Feb 27 08:52:22 2012]       [ Show / Hide ]
>>
>> run_in c:/Inetpub/wwwroot/ISB/data/Olaf/OR_8; c:\Inetpub\tpp-bin
>> \xinteract  -Now_8_1-3.pep.xml -p0.05 -l7 -OAgx -I1 -
>> OT2_Gundry_2012-02-21_OR-08_1.pep.xml -
>> OT2_Gundry_2012-02-21_OR-08_2.pep.xml -
>> OT2_Gundry_2012-02-21_OR-08_3.pep.xml
>>
>> c:\Inetpub\tpp-bin\xinteract (TPP v4.5 RAPTURE rev 2, Build
>> 201202031108 (MinGW))
>>
>> running: "C:/Inetpub/tpp-bin/InteractParser "ow_8_1-3.pep.xml" -L"7""
>> processed altogether 0 results
>> ERROR: output file created, c:/Inetpub/wwwroot/ISB/data/Olaf/OR_8/
>> ow_8_1-3.pep.xml, appears incomplete. Please check the input files for
>> completeness.
>>
>>
>> results written to file c:/Inetpub/wwwroot/ISB/data/Olaf/OR_8/
>> ow_8_1-3.pep.xml
>>
>> direct your browser to
>> http://localhost/ISB/data/Olaf/OR_8/ow_8_1-3.pep.shtml
>>
>>
>>
>> command completed in 0 sec
>>
>> running: "C:/Inetpub/tpp-bin/DatabaseParser "ow_8_1-3.pep.xml""
>> command completed in 0 sec
>>
>> No single database specified (by -D option or in .pep.xml file).
>> RefreshParser skipped.
>>
>> running: "C:/Inetpub/tpp-bin/PeptideProphetParser "ow_8_1-3.pep.xml"
>> MINPROB=0.05 ACCMASS GLYC EXCLUDE IGNORECHG=1 NEGGAMMA NONEGINIT
>> DECOYPROBS RT EXCLUDE MALDI LEAVE NEGGAMMA NONEGINIT DECOYPROBS RT
>> EXCLUDE MALDI LEAVE NEGGAMMA NONEGINIT DECOYPROBS RT EXCLUDE MALDI
>> LEAVE"
>> using Accurate Mass Bins
>> using RT
>> using N-glyc info
>> maldi mode
>> Ignoring charge 1+ spectra.
>> Using hardcoded values to initialize the distributions.
>> Using Gamma Distribution to model the negative hits.
>> MS Instrument info: Manufacturer: UNKNOWN, Model: UNKNOWN, Ionization:
>> UNKNOWN, Analyzer: UNKNOWN, Detector: UNKNOWN
>>
>> Error: file update failed (did not find closing tag "</
>> msms_pipeline_analysis>"), check file ow_8_1-3.pep.xml.tmp.a04004 for
>> completeness
>> error: no peptideprophet data written to file ow_8_1-3.pep.xml
>>
>> command "C:/Inetpub/tpp-bin/PeptideProphetParser "ow_8_1-3.pep.xml"
>> MINPROB=0.05 ACCMASS GLYC EXCLUDE IGNORECHG=1 NEGGAMMA NONEGINIT
>> DECOYPROBS RT EXCLUDE MALDI LEAVE NEGGAMMA NONEGINIT DECOYPROBS RT
>> EXCLUDE MALDI LEAVE NEGGAMMA NONEGINIT DECOYPROBS RT EXCLUDE MALDI
>> LEAVE" failed: Operation not permitted
>>
>> command "C:/Inetpub/tpp-bin/PeptideProphetParser "ow_8_1-3.pep.xml"
>> MINPROB=0.05 ACCMASS GLYC EXCLUDE IGNORECHG=1 NEGGAMMA NONEGINIT
>> DECOYPROBS RT EXCLUDE MALDI LEAVE NEGGAMMA NONEGINIT DECOYPROBS RT
>> EXCLUDE MALDI LEAVE NEGGAMMA NONEGINIT DECOYPROBS RT EXCLUDE MALDI
>> LEAVE" exited with non-zero exit code: 1
>> QUIT - the job is incomplete
>>
>> command "c:\Inetpub\tpp-bin\xinteract -Now_8_1-3.pep.xml -p0.05 -l7 -
>> OAgx -I1 -OT2_Gundry_2012-02-21_OR-08_1.pep.xml -
>> OT2_Gundry_2012-02-21_OR-08_2.pep.xml -
>> OT2_Gundry_2012-02-21_OR-08_3.pep.xml" failed: Operation not permitted
>>
>> Command FAILED
>>
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>

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