Hi Natalie,

Thanks for the info but my question is still not answered completely.

My question was "*HOW*" TPP does what it does. We all know it extracts more
info from results but I wanted to know HOW does it calculate the
statistically valid p-value (chance of match being incorrect given the null
hypothesis) if the search engine did i already?

If u know the database size, u can always calculate the p-value from e-value
and vice-versa. So, how does TPP fit in?

Lets focus on another very potent problem not much discussed in search
engines - In higher mass ranges there are very few candidates to accurately
calculate a p-value or e-value (or any statistical measure for that matter).
Unless u have a null model (a supposedly bell curve for random hits), u
cannot calculate a statistical confidence. And u dont have enough candidates
(lets assume just 5 candidates) to draw a distribution for Tandem to
calculate the e-value correctly.
Curve fitting will have drastic effect here.

How does TPP correct for it, if it does so?

Regards,

Amit Kumar Yadav
Senior Research Fellow (SRF-CSIR)
IGIB, New Delhi (India)

http://masswiz.igib.res.in




On Thu, Mar 11, 2010 at 11:55 AM, Natalie Tasman <
[email protected]> wrote:

> Good questions and worth explaining for those new to the field.  X!Tandem
> is a program which to assign peptide sequences ("ID"s) to ms/ms spectra.  We
> call this type of program a "search engine" (for "peptide ID search engine"
> or similar).  Other programs in this class are OMSSA, Sequest, Mascot, and
> others.  Each of these programs can be run on its own, and outputs a score
> for each "assigned" peptide.  This score reflects the search engine's
> estimation of how likely or confident that assignment is (I use those terms
> not necessarily as true stats meanings here.)
>
> So now you have input of peptide IDs and "scores".  The TPP tools (Pep and
> Prot Prophet) do two major things of interest.  One, they take the
> variously-derrived scores, compute additional information not necessarily
> accounted by the search engine (asking such questions as "how reasonable is
> this sequence, given its terminii, length, hydrophobicity, and so on), and
> combine these numbers to arrive at a statistically valid p value.  Two, the
> TPP tools do this for many supported search engines, which allows the
> possibility of comparing the peptide assignments to other results, i.e. in a
> publication context.  (For a more complex approach to the last point, look
> at the TPP's InterProphet tool.)
>
> Regarding FDR, I will leave that to someone else to answer in detail.
>
> Hope this helps,
> Natalie
>
> On 3/10/10 8:46 PM, Amit Kumar Yadav wrote:
>
>> Dear All,
>>
>> I was just browsing the gpm site and reading about tandem. It says
>> that peptideprophet and proteinprophet need not be used with X!Tandem.
>> Can someone explain (in simple terms please) how TPP uses tandem
>> result data to assign probabilities?
>>
>> Another naive question is about calculation of FDR from X!Tandem
>> results? How should it be done?
>>
>> For reference-
>> Quoting from http://www.thegpm.org/TANDEM/index.html "Unlike
>> some... ... Therefore, separate assembly and statistical analysis
>> software, e.g. PeptideProphet and ProteinProphet, do not need to be
>> used."
>>
>>
>>
>
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