Hi Jimmy,

thanks for the quick reply.

> > 1.) Is it possible to use 13C labeling with either XPress or
> > ASAPRatio? I've found only options for 15N, so is there any chance to
> > process 13C-labeled samples?
>
> You can't analyze 13C labeling now with Xpress.  I don't see 13C (nor
> 15N) support mentioned in the ASAPRatioPeptideParser usage statement so
> there's a decent chance you can't use ASAPRatio right now for 13C
> analysis either.

Okay, now I've also seen that 15N support is not mentioned in the
ASAPRatio usage statement. I was quite sure that I have read about 15N
support for ASAPRatio in the announcement of TPP4.2. Perhaps I was
just fantasizing.

>
> It was easy to extend the 15N option in Xpress to 13C labeling which I
> just did with the source files I have checked out.  But it might take
> awhile before these changes make it out.  I need to check in the mods,
> we'll need a small TPP GUI update, and the next TPP release will need to
> be released.

Oh, this would be great if your modifications were available. For the
moment I would also be happy if you could just check in the files so I
could get them and compile them here (or, if you don't mind, even mail
them to me. I have some people here urgently waiting for some
results...). I would also do some beta- (or alpha-) testing of the new
feature ;-)

>
> > 2.) How to perform the searches (Mascot in my case)? As far as I
> > understood I have to search each sample twice - once for the light and
> > once for the heavy peptides. That means to modify the masses and
> > modification files on the Mascot server for every heavy search, so it
> > is quite uncomfortable, especially if the Mascot server is shared with
> > a lot of other people... With version 2.2, Mascot offers an option for
> > metabolic labeling, resulting in the identification of light and heavy
> > peptides in one run. Is it possible to process such a combined result
> > file through the TPP?
>
> For current Xpress analysis of metabolic labeled samples, the searches
> do need to be performed separately.
>
> I don't know how the modification specification in pepXML handles
> metabolic labeling and if the Mascot to pepXML converter can correctly
> translate combined light/heavy metabolic search results to pep.xml
> files.  It's going to take time and effort from some volunteer to make
> it possible to process a combined result through the TPP.

Okay, that's what I expected. After doing some seperated searches now
I have the impression that the pepXML does not handle metabolic
labeling at all. For heavy peptides, the Spectrum view (plot-msms)
shows complete nonsense. Also the mass of modifications on heavy
peptides is wrong - methionine oxidation is recorded with a mass of
147 but should be 148 on an N15 peptide. Or have I done something
wrong with the Mascot search or conversion?

Regarding Samy's mail, I have also found that the XPressDisplay shows
only the light peptides. I think there is a bug in
XPressCGIProteinDisplay.cxx - double use of the int k in two nested
for loops. I can provide a fixed version if someone is interested ;-)

Best wishes
- Matthias
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