Peptide list does not show all peptides derived from protein of interest

CPAS Forum (Inactive)
Peptide list does not show all peptides derived from protein of interest anette.karle  2011-02-25 10:15
Status: Closed
 
Dear Labkey team

I have a case where a peptide can match several sequences in the database - however since I put it into the sample I am sure which one is the correct one. If I use the search function in Labkey and enter the protein name I will get a list of all runs in which peps of this protein were found. If I click on one of the blue numbers in the "group" column I get a list with all proteins which contain this peptide and also at the end of the page there is the full list of peptides. However in this list there is no possibility to filter for XCorr and DCn.

As an alternative, if I start from the dashboard instead, click on a run, filter for the protein of interest in the protein name column, I only get a very short list of peptides - those where I only have 1 Sequence hit.
Where have all the others gone? I assuem that it only shows the peps where the score for "my protein" was higher than for other proteins. Since I know that only my protein can be the tue one, I don't care if one other protein with the identical sequence stretch scored higher for whatever reason - I just need all peptides shown.

How can I have a list of all peptides from this protein (regardless of the rank) and at the same time filter for DCn and for charge dependend XCorr values?

I would be glad if you could help me with this as soon as possible.

Best regards
Anette
 
 
Brian Connolly responded:  2011-02-28 13:22
Anette,

I apologize for the delayed response to your question. In order to do what you ask you will need to use one of LabKey's legacy views. The first step will be to modify the existing view to add in the the DetaCN(DCn) and XCorr columns and then use that view while looking at the proteins in your run. The instructions are below:

1) Start from the dashboard and click on a run.
2) Create a new view
  a) In the webpart named "View", in the Grouping drop down select "Protein (Legacy)" and hit the "go" button next to the drop down
  b) You will now see a new button, in the View webpart, named "Pick Peptide Columns"
  c) Click on the button named "Pick Peptide Columns"
  d) In the text box for the "Current:" field append the following (without the quotes) ',DeltaCN, XCorr'
  f) Click on the "Pick Columns" button
  g) This will bring you back to the previous page
  h) Click on the "Save View" button, in the View webpart, and enter a name of your choosing for this new view.
3) In the View webpart, find the drop down that says " <Select a saved View> " and choose the view that you created in the previous step. Hit the "go" button next to the drop down
4) In the View webpart, in the Grouping drop down, select "Protein (Legacy)". Click on the checkbox for "Expanded" and hit the "go" button next to the drop down.


In the Proteins webpart, you will now see for each Protein found in the run, there is a list of peptides in a table. You should see that DCn and XCorr columns are now in this table(in the last 2 columns). If you click on the column headers in this table, you will notice that you can sort and filter the data using these.

Please note, that you might have a Peptide filter applied to your data which limits the peptides and proteins shown in the view. Any filters or sorts applied to this run, will be shown in the area above the "Proteins" webpart.

Lastly, in step 3 above if you choose

- "Protein (Legacy)" in the drop down, it will display protein information from the search engine
- "ProteinProphet (Legacy)" in the drop down, it will display protein information from the ProteinProphet analysis



* In our next version (11.1) we will be introducing a new workflow to make this much more simple.
 
anette.karle responded:  2011-03-17 04:53
Hi Brian,
thanks for your reply.

I went through all the steps mentioned above, but I still don't get the full list of peptides which match this protein. The issue is that I have several other proteins with partially identical sequence in the database. So as soon as one peptide derived from my protein (which I know is the correct hit) is also matching one of those other proteins - this peptide won't show up in the peptide list.

How can I have both: possibility to filter for charge dependent XCorr values and one DCn value and at the same time get the full list of peptides no matter how many other proteins in the database match as well?

Best regards
Anette