Hi!

During postprocessing of InterProphet results (pepXML), I've seen some
irregularities that I don't understand. It would be great if someone
could shed some light on them.

1. additional modifications in X! Tandem results:
X! Tandem seems to add some modifications to the search parameters
that were not explicitly specified. The relevant lines from the pepXML
are below (sub-elements of "search_summary"). What's the meaning of
those?

<aminoacid_modification aminoacid="C" massdiff="-17.0265"
mass="143.0041" variable="Y" symbol="^"/>
<!--X! Tandem n-terminal AA variable modification-->
<aminoacid_modification aminoacid="E" massdiff="-18.0106"
mass="111.0320" variable="Y" symbol="^"/>
<!--X! Tandem n-terminal AA variable modification-->
<aminoacid_modification aminoacid="Q" massdiff="-17.0265"
mass="111.0321" variable="Y" symbol="^"/>
<!--X! Tandem n-terminal AA variable modification-->


2. alternative proteins in OMSSA results:
OMSSA doesn't seem to handle peptides that occur in different proteins
in the database consistently. It may not list alternative proteins and
give different protein references for the same peptide (A), or it may
give different numbers of alternative proteins for the same peptide
(B). Examples below. Is there a way to make sense of that or is it a
bug?

A: (note different "protein" values)

<search_hit hit_rank="1" peptide="SVLTVLHQDWLNGK" peptide_prev_aa="V"
peptide_next_aa="E" protein="P01860" num_tot_proteins="3"
num_matched_ions="20" tot_num_ions="26"
calc_neutral_pep_mass="1608.862" massdiff="0.013" is_rejected="0"
protein_descr="[snip]" num_tol_term="1" num_missed_cleavages="0">
<search_score name="pvalue" value="0.000000000000005"/>
<search_score name="expect" value="0.000000000413624"/>

<search_hit hit_rank="1" peptide="SVLTVLHQDWLNGK" peptide_prev_aa="V"
peptide_next_aa="E" protein="P01861" num_tot_proteins="3"
num_matched_ions="19" tot_num_ions="26"
calc_neutral_pep_mass="1608.862" massdiff="0.006999999999915"
is_rejected="0" protein_descr="[snip]" num_tol_term="1"
num_missed_cleavages="0">
<search_score name="pvalue" value="0.000000000003367"/>
<search_score name="expect" value="0.000000279733183"/>


B: (note different number of alternative proteins)

<search_hit hit_rank="1" peptide="FLEQQNQVLQTK" peptide_prev_aa="R"
peptide_next_aa="W" protein="P04264_CHAIN_0" num_tot_proteins="3"
num_matched_ions="20" tot_num_ions="22"
calc_neutral_pep_mass="1474.78" massdiff="0.007000000000035"
is_rejected="0" protein_descr="[snip]" num_tol_term="2"
num_missed_cleavages="0">
<alternative_protein protein="P35908" protein_descr="[snip]"
num_tol_term="2" peptide_prev_aa="R" peptide_next_aa="W"/>
<alternative_protein protein="P04264" protein_descr="[snip]"
num_tol_term="2" peptide_prev_aa="R" peptide_next_aa="W"/>
<search_score name="pvalue" value="0.000000000000005"/>
<search_score name="expect" value="0.000000000451822"/>

<search_hit hit_rank="1" peptide="FLEQQNQVLQTK" peptide_prev_aa="V"
peptide_next_aa="W" protein="Q7Z794" num_tot_proteins="4"
num_matched_ions="19" tot_num_ions="22"
calc_neutral_pep_mass="1474.78" massdiff="0.007000000000035"
is_rejected="0" protein_descr=[snip]" num_tol_term="1"
num_missed_cleavages="0">
<alternative_protein protein="P35908" protein_descr="[snip]"
num_tol_term="2" peptide_prev_aa="R" peptide_next_aa="W"/>
<alternative_protein protein="P04264" protein_descr="[snip]"
num_tol_term="2" peptide_prev_aa="R" peptide_next_aa="W"/>
<alternative_protein protein="P04264_CHAIN_0" protein_descr="[snip]"
num_tol_term="2" peptide_prev_aa="R" peptide_next_aa="W"/>
<search_score name="pvalue" value="0.000000000000001"/>
<search_score name="expect" value="0.000000000071547"/>


Thanks in advance

Hendrik

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