Hi all,

I am trying to run TPP with the my command 

xinteract -eN -p0 -dDECOY -OpPdAMN *files

for my comet searched files as my search enzyme is Formic Acid. However TPP 
seems like it is unable to complete the analysis for generating the Peptide 
Prophet Probabilities and thus cannot go through Protein Prophet analysis.

Below is the log file for the TPP's analysis. Is there any way that I can 
send my files through TPP and have a complete analysis?

xinteract (TPP v4.8.0 PHILAE, Build 201710121955-exported (CentOS-x86_64))

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/InteractParser 
'interact.pep.xml' '1.pep.xml' -L'7' -E'nonspecific'"
 file 1: 1.pep.xml
SUCCESS: CORRECTED data file /home/rostlab/annieha/BSA/mzML/1.mzML in 
msms_run_summary tag ...
 processed altogether 6293 results
INFO: Results written to file: 
/home/rostlab/annieha/BSA/comet/interact.pep.xml
command completed in 2 sec 

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/DatabaseParser 
'interact.pep.xml'"
command completed in 0 sec 

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/RefreshParser 
'interact.pep.xml' '/home/rostlab/annieha/BSA/BSA_target_decoy.fasta'"
  - Building Commentz-Walter keyword tree...  - Searching the tree...
  - Linking duplicate entries...  - Printing results...

command completed in 0 sec 

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/PeptideProphetParser 
'interact.pep.xml' MINPROB=0 DECOY=DECOY DECOYPROBS NONPARAM ACCMASS NONMC 
NONTT"
using Accurate Mass Bins
Using Decoy Label "DECOY".
Decoy Probabilities will be reported.
Not using ntt model
Not using nmc model
Using non-parametric distributions
 (Comet) (minprob 0)
adding Accurate Mass mixture distr
init with Comet nonspecific 
MS Instrument info: Manufacturer: unknown, Model: unknown, Ionization: 
UNKNOWN, Analyzer: UNKNOWN, Detector: UNKNOWN

 PeptideProphet  (TPP v4.8.0 PHILAE, Build 201710121955-exported 
(CentOS-x86_64)) AKeller@ISB
 read in 0 1+, 29 2+, 20 3+, 16 4+, 65 5+, 44 6+, and 0 7+ spectra.
Initialising statistical models ...
Found 87 Decoys, and 87 Non-Decoys
Iterations: .........10.........20.
WARNING: Mixture model quality test failed for charge (1+).
WARNING: Mixture model quality test failed for charge (2+).
WARNING: Mixture model quality test failed for charge (3+).
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+).
model complete after 22 iterations
command completed in 1 sec 

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl -i 
interact.pep.xml -d "DECOY""
Analyzing interact.pep.xml ...
Reading Accurate Mass Model model +1 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 217.
Reading Accurate Mass Model model +2 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 374.
Reading Accurate Mass Model model +3 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 531.
Reading Accurate Mass Model model +4 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 688.
Reading Accurate Mass Model model +5 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 845.
Reading Accurate Mass Model model +6 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 1002.
Reading Accurate Mass Model model +7 ...
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 311, <INFILE> 
line 1159.
Parsing search results "/home/rostlab/annieha/BSA/mzML/1 (Comet)"...
  => Found 174 hits. (87 decoys, 0 excluded)
  => Total so far: 174 hits. (87 decoys, 0 excluded)
Using decoy ratio of 0.5 for PeptideProphet.
Can't exec "/usr/bin/gnuplot": No such file or directory at 
/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProphetModels.pl line 903.
command completed in 62 sec 

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/cgi-bin/PepXMLViewer.cgi -I 
/home/rostlab/annieha/BSA/comet/interact.pep.xml"
command completed in 1 sec 

running: "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProteinProphet 
'interact.pep.xml' 'interact.prot.xml' XML"
ProteinProphet (C++) by Insilicos LLC and LabKey Software, after the 
original Perl by A. Keller (TPP v4.8.0 PHILAE, Build 201710121955-exported 
(CentOS-x86_64))
 (xml input) (no FPKM) (report Protein Length) (using degen pep info)
. . . reading in /home/rostlab/annieha/BSA/comet/interact.pep.xml. . .
. . . read in 0 1+, 0 2+, 0 3+, 0 4+, 0 5+, 0 6+, 0 7+ spectra with min 
prob 0.05
WARNING: no data - output file will be empty

command "/home/rostlab/hrost/bin/tpp/tpp4.8/bin/ProteinProphet 
'interact.pep.xml' 'interact.prot.xml' XML" exited with non-zero exit code: 
256
QUIT - the job is incomplete


Thanks,
Annie Ha


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