Dear Jimmy,

thanks for your quick answer. I will for sure look into the raw data
and hope to see a trend which is the reason for this effect.

What I forgot to mention in the post before, is that I post filtered
the results with a very stringent PPS cutoff of 0.99. So the reason
cannot be false positives.

Lukas


On Sep 15, 1:19 am, Jimmy Eng <[email protected]> wrote:
> Lukas,
>
> These tools could definitely be optimized and improved, especially for
> high res data that didn't really exist when they were first developed.
>  That said, with respect to question 1, you're using two imperfect
> tools on presumably large datasets so it's not surprising to see some
> uncorrelated results.  If I were you, I would look at the extracted
> chromatograms of these peptides to see how the two tools determined
> the respective light and heavy areas used to calculate the ratios.
> One (or both) likely failed for one reason or another which is one of
> the unfortunate reasons why the not-very-user-friendly GUIs exist to
> allow the researchers to look at and fix/edit/discard any particular
> calculated ratio.  I know you don't want to look at chromatograms for
> a quarter of your identifications but that's what you probably have to
> do at this point.  Or throw them out.  Or stick with a single tool and
> pretend the other didn't exist that gives conflicting results.
>
> I don't know the answers to your other two questions, not that I
> really had an answer to question 1, so hopefully someone else can
> address those.
>
> - Jimmy
>
>
>
> On Tue, Sep 14, 2010 at 12:42 PM, lukas <[email protected]> wrote:
> > Dear list,
>
> > I have three question regarding ASAP and XPRESS:
> > 1)
> > I analyzed a data set generated from a SILAC sample on an Orbitrap.
> > The data was searched using Sequest with variable modifications on R
> > and K. I used ASAP and XPRESS for quantitaton with the following
> > command (TPP v4.3 JETSTREAM rev 1, Build 201004270833 (linux)):
> > xinteract -dreverse_ -p0.8 -OAlpd -X-m0.1-nK,8.014199-nR,10.008269 -A-
> > Z-r0.1-F-C-lKR-mK136.10916R166.109379 *.pep.xml
>
> > When I make the correlation between log2("asap mean") and
> > log2("xpress") I can see a large part of the data points nicely
> > correlating but also many data points (roughly a quarter) having close
> > to zero regulation by XPRESS but non-zero regulation by ASAP.
> > I wonder whether I chose any wrong options or what the reason for this
> > effect could be? Any hint would be greatly appreciated.
>
> > 2)
> > ASAP threw the warning message:
> > WARNING: Found more than one variable mod on 'K'. Please make sure to
> > specify a heavy mass for this residue.
> > WARNING: Found more than one variable mod on 'R'. Please make sure to
> > specify a heavy mass for this residue.
>
> > Could it be that the reason for this are peptides that were identified
> > with one light and one heavy K/R and are most likely false positives?
>
> > 3)
> > Where can I read about the difference between "asap mean" and
> > "asapratio_HL"?
>
> > Thanks for any help,
> > Lukas
>
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