Hi Brian,

I just confirmed in the code that it is counted here when the weight of the
peptide is minimum 0.5 and the probability minimum 0.2.

Cheers,
-David

On Wed, Jun 28, 2023 at 9:49 AM Hampton, Brian <[email protected]>
wrote:

> Hi David, thanks for the quick reply.
>
> I think I see now.  When there is some ambiguity which prevents
> determining a precise peptide sequence, for example incomplete fragment ion
> series, or isobaric AA assignments etc. then there must be a probability
> calculation performed, the result of which attempts to assign this peptide
> to the more likely protein candidate.  The result of which is recorded in
> the "total num peptides" value.  In the case of a tie, is the value added
> to all possible protein hits or not used?
>
> Thanks a bunch!
>
> Brian
> ------------------------------
> *From:* 'David Shteynberg' via spctools-discuss <
> [email protected]>
> *Sent:* Wednesday, June 28, 2023 12:37 PM
> *To:* [email protected] <[email protected]
> >
> *Subject:* Re: [spctools-discuss] ProteinProphet output number of unique
> peptides vs total number of peptides
>
> Hello Brian,
>
> Thanks for the discussion.  The way this is possible is because there are
> degenerate peptides that map to multiple proteins, in which case only those
> that have sufficient weight will be counted as "is contributing evidence"
> peptides.  Only those PSM  instances that are contributing evidence are
> counted in the "tot num peps" value.   Please let me know if further
> clarification would be helpful or if you have other questions.
>
> Cheers,
> -David
>
> On Wed, Jun 28, 2023 at 9:27 AM Hampton, Brian <[email protected]>
> wrote:
>
> Hello David,
>
> I have difficulty understanding this.  I had always operated under the
> assumption that the "num unique peps" would be less than or equal to the
> "tot num peps" - because - I thought "tot num peps" = "num unique peps" +
> number of (shared) peptides also mapping to given protein.  Is this not the
> case and if so how would "tot num peps" ever be less than "num unique peps"?
>
> Best regards,
> Brian
>
>
>
> Brian Hampton
> Proteomics Core Lab
>
> Center for Vascular and Inflammatory Diseases
>
> University of Maryland School of Medicine
>
> BioPark One Rm 307
>
> 800 West Baltimore Street
>
> Baltimore, MD. 21201
>
> (410)706-8207
> ------------------------------
> *From:* 'David Shteynberg' via spctools-discuss <
> [email protected]>
> *Sent:* Wednesday, June 28, 2023 11:33 AM
> *To:* [email protected] <[email protected]
> >
> *Subject:* Re: [spctools-discuss] ProteinProphet output number of unique
> peptides vs total number of peptides
>
> Hi Murielle,
>
> Sorry this is a little confusing.  Column "num unique peps" represents
> total number of unique peptides mapping to the protein.  The "tot num peps"
> counts the PSMs that are "contributing evidence" (last column) to the
> protein, sometimes this number can be lower because not all peptides
> mapping are contributing evidence and spectral counts can be low per
> peptide.
>
> The answer to your second question is *yes*, the "tot num peps" is the
> spectral count for "is contributing evidence" peptides.
>
> Cheers,
> -David
>
>
> On Wed, Jun 28, 2023 at 7:27 AM [email protected] <
> [email protected]> wrote:
>
> Hi David,
>
> Thank you for such a quick reply!
>
> I used tpp version 6.2.0 run via a singularity image and I produced the
> table by including the EXCELXX option in proteinprophet.
>
> Cheers,
>
> Murielle
>
> On Wednesday, June 28, 2023 at 12:44:03 AM UTC+2 David Shteynberg wrote:
>
> Hi Murielle,
>
> Thanks for using the TPP and submitting your question here.   When I try
> the latest version of the TPP and run ProteinProphet through there, the
> columns exported in my tab separated file from ProteinProphet are different
> from yours.  Can you please provide a bit more information about how you
> generated this file and which version of the software you used?
>
> Cheers,
> -David
>
> On Tue, Jun 27, 2023 at 7:10 AM [email protected] <[email protected]>
> wrote:
>
> Hi all,
>
> I have two questions concerning the output of ProteinProphet :
>
> 1) Why do I have some entries for which the total number of peptides is
> lower than the number of unique peptides?  (see columns 5 and 6 below, an
> extract of my output table)
>
> [image: protein prophet output.jpeg]
> 2) Does the column "total number of peptides" correspond to spectral
> counts?
>
> Thank you very much ,
>
> Murielle
>
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