Hi,

I am trying to run peptideprophet on Phosphoproteomics Data from Lumos. 

I am using the following command line on comet search results (note: Tandem 
kscore went through successfully)


*xinteract -OARPwd -dDECOY_  
-N/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml
 *.comet.pep.xml*

*This is my output log:*

xinteract (TPP v5.2.0 Flammagenitus, Build 201903181054-exported 
(Linux-x86_64))

running: "/hpc/apps/TPP/5.2.0/bin/DatabaseParser 
'/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml'"
command completed in 24 sec 

running: "/hpc/apps/TPP/5.2.0/bin/RefreshParser 
'/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml'
 
'/scratch/soetkampd/iphronesis/reference_files/Databases/Uniprot_rat_8060_20190321_DECOY.fasta'"

  - Searching the tree...
  - Linking duplicate entries...
  - Printing results...

command completed in 264 sec 

running: "/hpc/apps/TPP/5.2.0/bin/PeptideProphetParser 
'/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml'
 
ACCMASS RT NONPARAM INSTRWARN DECOYPROBS DECOY=DECOY_"
using Accurate Mass Bins
using RT
Using no error on different instrument types.
Using Decoy Label "DECOY_".
Decoy Probabilities will be reported.
Using non-parametric distributions
 (Comet)
adding ACCMASS mixture distribution
adding Retention Time mixture distr
init with Comet trypsin 
MS Instrument info: Manufacturer: Thermo Scientific, Model: Orbitrap 
Fusion, Ionization: UNKNOWN, Analyzer: UNKNOWN, Detector: UNKNOWN

INFO: Processing standard MixtureModel ... 
Initialising statistical models ...
Iterations: .........10.........20.....Estimating Retention Time Model ... 
please wait ... 

model complete after 26 iterations
command completed in 5180 sec 

running: "/hpc/apps/TPP/5.2.0/bin/ProphetModels.pl -i 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml
 
-d "DECOY_""
Analyzing 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml
 
...
Reading Accurate Mass Model model +1 ...
Reading kernel density RTCalc RT [RT] model +1 ...
Reading Accurate Mass Model model +2 ...
Reading kernel density RTCalc RT [RT] model +2 ...
Reading Accurate Mass Model model +3 ...
Reading kernel density RTCalc RT [RT] model +3 ...
Reading Accurate Mass Model model +4 ...
Reading kernel density RTCalc RT [RT] model +4 ...
Reading Accurate Mass Model model +5 ...
Reading kernel density RTCalc RT [RT] model +5 ...
Reading Accurate Mass Model model +6 ...
Reading kernel density RTCalc RT [RT] model +6 ...
Reading Accurate Mass Model model +7 ...
Reading kernel density RTCalc RT [RT] model +7 ...
Parsing search results 
"/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_54_Phospho_centroid
 
(Comet)"...
  => Found 0 hits. (0 decoys, 0 excluded)
  => Total so far: 0 hits. (0 decoys, 0 excluded)
....
....
....
Parsing search results 
"/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_01_Phospho_centroid
 
(Comet)"...
  => Found 0 hits. (0 decoys, 0 excluded)
  => Total so far: 0 hits. (0 decoys, 0 excluded)
command completed in 5 sec 

running: "/hpc/apps/TPP/5.2.0/cgi-bin/PepXMLViewer.cgi -I 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml"

*command "/hpc/apps/TPP/5.2.0/cgi-bin/PepXMLViewer.cgi -I 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.comet.interact.pep.xml"
 
exited with non-zero exit code: 139*
*QUIT - the job is incomplete*


*This is my error log:*

INFO: Results written to file: 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/peptideprophet/2018.kscore.interact.pep.xml
  - Building Commentz-Walter keyword tree... PeptideProphet  (TPP v5.2.0 
Flammagenitus, Build 201903181054-exported (Linux-x86_64)) AKeller@ISB
 read in 0 1+, 495215 2+, 990646 3+, 429079 4+, 105824 5+, 0 6+, and 0 7+ 
spectra.
Found 971133 Decoys, and 1049631 Non-Decoys
Run Index: 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_30_Phospho_centroid,
 
slope=0.510115, intercept=665.195, r_sq=0.633259
Run Index: 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_09_Phospho_centroid,
 
slope=0.420776, intercept=663.458, r_sq=0.679369
WARNING: Not enough correlation r_sq=0.268561 between theoretical RT values 
and scan numbers in run index 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_55_Phospho_centroid.
 
RT Model has been disabled.
....
....
....
WARNING: Not enough correlation r_sq=0.296415 between theoretical RT values 
and scan numbers in run index 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_14_Phospho_centroid.
 
RT Model has been disabled.
Run Index: 
/scratch/soetkampd/iphronesis/platform_workspace/jobs/SJOB2018/msconvert/DS_190126_DDA_H9C2_14_Phospho_centroid,
 
slope=0.425669, intercept=394.448, r_sq=0.296415
WARNING: Mixture model quality test failed for charge (1+).
WARNING: Mixture model quality test failed for charge (4+).
WARNING: Mixture model quality test failed for charge (5+).
WARNING: Mixture model quality test failed for charge (6+).
WARNING: Mixture model quality test failed for charge (7+).

*I am wondering if there is something i might have missed in my 
command-line (specifically needed for Phospho data) or if this is a memory 
issue since i do have 60 DDA runs. Any advice on how to debug this will be 
highly appreciated.*
*I am happy to make these files available via Box if needed.*

Regards,

Elva

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