Made a mess of the example as Sequest used chymotrypsin while the TPP
pepXML creation process was set to trypsin. The following is a better
example:

Sequest output file:

 10HNE-1ACO-Trypsin4-Orbit1.8062.8062.2.out
 SEQUEST v.28 (rev. 13), (c) 1998-2007
 Molecular Biotechnology, Univ. of Washington, J.Eng/S.Morgan/J.Yates
 Licensed to Thermo Fisher Scientific Inc.
 02/22/2012, 07:00 PM, 0.3 sec. on VCU-D409FC038FB
 (M+H)+ mass = 1745.88017 ~ 0.0262 (+2), fragment tol = 0.0100, MONO/
MONO
 total inten = 8137.7, lowest Sp = 1.8, # matched peptides = 2361
 # amino acids = 267050, # proteins = 49378, C:\Xcalibur\database
\Aconitase_Pig_RefSeq_20120219_GPM_cRAP_DECOY.fasta
 ion series nABY ABCDVWXYZ: 0 1 1 0.0 1.0 0.0 0.0 0.0 0.0 0.0 1.0 0.0
 display top 10/1, ion % = 0.0, CODE = 001050
 (M* +15.99492) (C# +101.10922) (H@ +158.13068) (K^ +158.13068) (K~
+140.12012) (nt] -17.02655) C=160.03064  Enzyme:Trypsin(KR/P) (3)

  #   Rank/Sp      Id#     (M+H)+    deltCn   XCorr    Sp     Sf
Ions  Reference                            Peptide
 ---  --------  --------  --------   ------  ------   -----  ----
----  ---------                            -------
  1.   1 /  1          1 1745.87448  0.0000  3.4809   610.9  0.93
15/30  gi|47522738|ref|NP_999119.1|         K.Q]GLLPLTFADPADYNK.I
  2.   2 /  4      24102 1745.87448  0.7340  0.9259    33.8  0.00
4/26  gi|47523048|ref|NP_999285.1|         K.K]VVYKQYQDESFTK.R
  3.   3 / 37      17482 1745.89245  0.7641  0.8211    12.5  0.00
3/24  gi|335297918|ref|XP_003131644.2|     R.FRK^DMM*KNASESK.L
  4.   4 /  5      29008 1745.85672  0.7694  0.8028    32.4  0.00
4/26  decoy_4319                       +1  R.LFCIYSTLHESFTK.L
  5.   5 / 15      42361 1745.89694  0.7725  0.7919    20.0  0.00
3/28  decoy_17672                          K.EPVSSGLPNRQWTFK.S
  6.   6 /  8        401 1745.90434  0.7808  0.7630    26.2  0.00
3/24  gi|358679347|ref|NP_001240636.1|     K.E]HVK~WVHAEM*VNK.N
  7.   7 / 62       6413 1745.86661  0.7821  0.7584     8.3  0.00
2/28  gi|311250321|ref|XP_003124098.1|     R.YEVC#LAGGSGLDEFK.F
  8.   8 / 77      21448 1745.89561  0.7843  0.7507     5.8  0.00
2/28  gi|311259750|ref|XP_003128242.1|     R.Y]GSH@DGGSAAPDGK^K.K
  9.   9 /  7       6126 1745.86208  0.7875  0.7395    28.1  0.00
4/22  gi|311252102|ref|XP_003124922.1|     R.KWPRQYEWFFMK.L
 10.  10 / 77      10016 1745.85607  0.7887  0.7356     5.8  0.00
2/26  gi|350592534|ref|XP_001924671.3|     R.VGEYKKM*QQM*DTKK.L

  1.          1  gi|47522738|ref|NP_999119.1| aconitate hydratase,
mitochondrial precursor, minus transit peptide [Sus scrofa]

TPP pepXML result:

<?xml version="1.0" encoding="UTF-8"?>
<?xml-stylesheet type="text/xsl" href="http://regis-
web.systemsbiology.net/pepXML_std.xsl"?>
<msms_pipeline_analysis date="2012-02-23T16:10:09" xmlns="http://regis-
web.systemsbiology.net/pepXML" xmlns:xsi="http://www.w3.org/2001/
XMLSchema-instance" xsi:schemaLocation="http://regis-
web.systemsbiology.net/pepXML C:\Inetpub\wwwrootpepXML_v117.xsd"
summary_xml="c:/Inetpub/wwwroot/ISB/data/extension/N_Term_Example/
10HNE-1ACO-Trypsin4-Orbit1.pep.xml">
<msms_run_summary base_name="c:/Inetpub/wwwroot/ISB/data/extension/
N_Term_Example/10HNE-1ACO-Trypsin4-Orbit1" raw_data_type="raw"
raw_data="(null)">
<sample_enzyme name="trypsin">
<specificity cut="KR" no_cut="P" sense="C"/>
</sample_enzyme>
<search_summary base_name="c:/Inetpub/wwwroot/ISB/data/extension/
N_Term_Example/10HNE-1ACO-Trypsin4-Orbit1" search_engine="SEQUEST"
precursor_mass_type="monoisotopic" fragment_mass_type="monoisotopic"
out_data_type="out" out_data=".tgz" search_id="1">
<search_database local_path="C:\Inetpub\wwwroot\ISB\data\dbase
\Aconitase_Pig_RefSeq_20120219_GPM_cRAP_DECOY.fasta" type="AA"/>
<aminoacid_modification aminoacid="M" massdiff="15.9949"
mass="147.0354" variable="Y" symbol="*"/>
<aminoacid_modification aminoacid="C" massdiff="101.1092"
mass="261.1399" variable="Y" symbol="#"/>
<aminoacid_modification aminoacid="H" massdiff="158.1307"
mass="295.1896" variable="Y" symbol="@"/>
<aminoacid_modification aminoacid="K" massdiff="158.1307"
mass="286.2256" variable="Y" symbol="^"/>
<aminoacid_modification aminoacid="K" massdiff="140.1201"
mass="268.2151" variable="Y" symbol="~"/>
<aminoacid_modification aminoacid="C" massdiff="57.0215"
mass="160.0306" variable="N"/>
<terminal_modification terminus="n" massdiff="-17.0265"
mass="-16.0187" variable="Y" symbol="]" protein_terminus="N"/>
<parameter name="peptide_mass_tol" value="15.000"/>
<parameter name="fragment_ion_tol" value="0.010"/>
<parameter name="ion_series" value=" 0 1 1 0.0 1.0 0.0 0.0 0.0 0.0 0.0
1.0 0.0"/>
<parameter name="max_num_differential_AA_per_mod" value="4"/>
<parameter name="nucleotide_reading_frame" value="0"/>
<parameter name="num_output_lines" value="10"/>
<parameter name="remove_precursor_peak" value="0"/>
<parameter name="ion_cutoff_percentage" value="0.0"/>
<parameter name="match_peak_count" value="0"/>
<parameter name="match_peak_allowed_error" value="1"/>
<parameter name="match_peak_tolerance" value="1.0"/>
<parameter name="protein_mass_filter" value="0 0"/>
<parameter name="sequence_header_filter" value=""/>
</search_summary>
  <spectrum_query spectrum="10HNE-1ACO-Trypsin4-Orbit1.8062.8062.2"
start_scan="8062" end_scan="8062" precursor_neutral_mass="1744.8729"
assumed_charge="2" index="1">
    <search_result>
      <search_hit hit_rank="1" peptide="QGLLPLTFADPADYNK"
peptide_prev_aa="K" peptide_next_aa="I" protein="gi|47522738|ref|
NP_999119.1|" num_tot_proteins="1" num_matched_ions="15"
tot_num_ions="30" calc_neutral_pep_mass="1744.8672"
massdiff="+0.005690" num_tol_term="2" num_missed_cleavages="0"
is_rejected="0">
        <search_score name="xcorr" value="3.481"/>
        <search_score name="deltacn" value="0.734"/>
        <search_score name="deltacnstar" value="0.000"/>
        <search_score name="spscore" value="610.9000"/>
        <search_score name="sprank" value="1"/>
      </search_hit>
    </search_result>
  </spectrum_query>
</msms_run_summary>
</msms_pipeline_analysis>

David Simpson
Virginia Commonwealth University

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