Hi all,

I am running PeptideProphet, iProphet and ProteinProphet (TPP 4.6.3) on Q 
Exactive data searched with Comet, Myrimatch and OMSSA. I wondered if the 
NSP model should be disabled in ProteinProphet when it is enabled in 
iProphet? I got confused because it seems Petunia enables the NSP model 
both in iprophet and proteinprophet by default (ie. when xinteract runs 
with the -ip option).

Another question is that when I compare decoy estimated protein FDRs to 
ProteinProphet modelled FDRs, ProteinProphet seems a bit optimistic (decoy 
based FDR of 0.1% corresponds to ~0.02% model FDR). This is with NSP 
enabled in iProphet and disabled in ProteinProphet. How should I deal with 
discrepancy, ie. should I take the decoy or probability based FDR to select 
a probability cutoff?

I have attached some examples for a search with myrimatch only. These are 
the commands I used to generate the graphs:

xinteract -Nmyrimatch.pep.xml -OAP -p0 -a%ExperimentFolder% -dDECOY0 
-E%ExperimentTag% *.pep.xml
InterProphetParser myrimatch.pep.xml myrimatch.ipro.pep.xml
ProphetModels.pl -i myrimatch.ipro.pep.xml -k -r 0.25 -d "DECOY1"
ProteinProphet myrimatch.ipro.pep.xml myrimatch.prot.xml IPROPHET NONSP
ProtProphModels.pl -k -r 0.25 -d DECOY1 -i myrimatch.prot.xml

The graphs are:

myrimatch_all.ipro.pep_FDR_10pc: PeptideProphet/iProphet decoy vs model 
FDR, all models enabled
myrimatch_nonsp.ipro.pep_FDR_10pc: PeptideProphet/iProphet decoy vs model 
FDR, NSP model disabled in iProphet
myrimatch_nonsp.prot_FDR_5pc: ProteinProphet decoy vs model FDR, NSP model 
disabled in ProteinProphet
myrimatch_all.prot_FDR_5pc: ProteinProphet decoy vs model FDR, NSP model 
enabled in iProphet and ProteinProphet

Thanks in advance!

Kind regards,

Rene


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