QuickMod help


Version 27/07/2010


What is QuickMod?


QuickMod is a spectral library search based MSMS data analysis tool, designed to identify modified peptides.

A spectral library is a database of already identified MSMS spectra. Typically a peptide entry in the library is represented by a consensus spectrum; an averaged spectrum compiled from multiple, same precursor charge state, spectra representing the same peptide species.

The QuickMod algorithm assumes that the precursor mass difference between a query spectrum and a candidate library spectrum can be explained by a modification. Based on this assumption the two spectra are aligned and a list of similarity measures calculated. In a second step the most likely attachment position of the modification is determined.

The QuickMod scoring scheme was developed by carefully examining and combining various peak intensity and peak coverage based scoring features. Using a similar approach proposed for the Percolator algorithm (Käll et al., Nature Methods 2007, The optimal linear combination of the list of scoring features is derived using a Support Vector Machine classification algorithm).

QuickMod provides efficient means to evaluate the quality of a MS/MS dataset by revealing modifications induced by sample handling and preparation such as oxidation, pyro-Glu and salt adducts. More importantly, the search algorithm is capable of identifying known PTMs while suggesting a list of previously unknown or unexpected PTMs, to be verified in targeted experiments.



Input parameters


Data file

Currently, QuickMod only accepts spectra in Mascot Generic Format (MGF).
Please have a look here for a general description of this format.

In the « Data file content » area, one can directly paste a list of spectra in MGF.

In a test purpose, one can also load an example spectrum corresponding to this peptide/modification: M({15.999})PcAEDYLSVVLNQLcVLHEK


Spectral libraries

The spectral libraries available in this web-interface version of QuickMod were built by S. Stein at NIST; more information and related software can be found at peptide.nist.gov.

LibrarySpectraRelease dateDetailed content
Human Ion-Trap345 48914/01/2010http://peptide.nist.gov/info/human.php
Human QTOF14 82702/03/2010http://peptide.nist.gov/info/human_qtof.php
Human Serum Albumin, Ion-Trap2 30903/08/2009http://peptide.nist.gov/info/hsa.php
Yeast Ion-Trap90 50719/10/2009http://peptide.nist.gov/info/yeast.php
Yeast QTOF2 41825/11/2009http://peptide.nist.gov/info/yeast_qtof.php


Other general parameters


Precursor mass tolerance

Allowed deviation (in m/z) between the precursor mass from the spectrum and the mass of candidate peptides from the database.

Fragment mass tolerance

Specify an error tolerance (in m/z) for MS/MS fragment ion mass values.

Modification mass tolerance

Additional allowed mass tolerance for modifications.


Output parameters


Number of candidates per spectrum

Number of possible results proposed by QuickMod for each spectrum.

Score threshold

Only results above this value will be displayed in the output. Recommended value is 0.6. More details on the score can be found here.

Your E-mail adress

If filled, the result will be sent to this e-mail adress.


Output


Peptide

Example: M({15.999})PcAEDYLSVVLNQLcVLHEK

({15.999}) : indicates a modification of approximatively 16 Da on Methionine
c (lower case) : the query spectrum was matched to a library spectrum representing a peptide with a Cys-Cam modification.


DP or Dot Product score

The current web-interface version of QuickMod calculates the similarity between a query spectrum and its spectral library candidates using a cross-correlation like scoring function. Similarity scores range from 0-1;

Approximate score interpretation

dp = 0 - no similarity

dp < 0.6 - unlikely match

dp 0.6 > dp > 0.8 - good match

dp >0.8 - excellent match

dp = 1 - perfect match



Any further question can be adressed to the ExPASy helpdesk.