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<HTML>
<HEAD>
<TITLE>
EMBOSS: digest
</TITLE>
</HEAD>
<BODY BGCOLOR="#FFFFFF" text="#000000">
<table align=center border=0 cellspacing=0 cellpadding=0>
<tr><td valign=top>
<A HREF="/" ONMOUSEOVER="self.status='Go to the EMBOSS home page';return true"><img border=0 src="emboss_icon.jpg" alt="" width=150 height=48></a>
</td>
<td align=left valign=middle>
<b><font size="+6">
digest
</font></b>
</td></tr>
</table>
<br>
<p>
<H2>
Function
</H2>
Protein proteolytic enzyme or reagent cleavage digest
<H2>
Description
</H2>
<b>digest</b> finds the positions where a specified proteolytic enzyme
or reagent might cut a peptide sequence.
<p>
This programs allows you to input a protein sequence and to specify one
proteolytic agent from a list. It will then output a file containing
the positions where the agent cuts, together with the peptides produced.
<p>
Trypsin will not normally cut after a K if it is followed by (e.g.)
another K or a P. Specifying the qualifier <b>-unfavoured</b> shows
those cuts. as well as the favoured ones.
<p>
If you wish to emulate a partial digestion, then using the
<b>-overlap</b> qualifier will display the results from a digest in
which all cut sites are used and in which one site at a time is not cut.
Thus the resulting peptide fragments from the cut sites numbered 1, 2,
3, 4 etc. are shown, plus the fragments produced by cutting at the
sites 1..3, 2..4, etc.
<p>
i.e. fragments containing one potential cut site are also shown.
<p>
If you wish to emulate a <b>very</b> partial digestion (!) then using
the <b>-allpartials</b> qualifier will do what <b>-overlap</b> did, but
also show all possible fragments from using all possible combinations of
cut sites. For example, fragments from the cut sites numbered 1, 2, 3,
4, 5 and from the fragments produced by cutting between sites 1..3,
1..4, 1..5, 2..4, 2..5, 3..5, etc.
<p>
i.e. fragments containing one or more potential cut site are also shown.
<p>
If the boolean -ragging is specified then terminal break-up
fragments are also shown; this can be useful for mass spectrometry.
Ragging is further controlled by the -termini
option which allows you to select removal of residues from
the N and/or C termini.
<H2>
Usage
</H2>
<b>Here is a sample session with digest</b>
<p>
<p>
<table width="90%"><tr><td bgcolor="#CCFFFF"><pre>
% <b>digest </b>
Protein proteolytic enzyme or reagent cleavage digest
Input protein sequence(s): <b>tsw:opsd_human</b>
Enzymes and Reagents
1 : Trypsin
2 : Lys-C
3 : Arg-C
4 : Asp-N
5 : V8-bicarb
6 : V8-phosph
7 : Chymotrypsin
8 : CNBr
Select number [1]: <b></b>
Output report [opsd_human.digest]: <b></b>
</pre></td></tr></table><p>
<p>
<a href="#input.1">Go to the input files for this example</a><br><a href="#output.1">Go to the output files for this example</a><p><p>
<H2>
Command line arguments
</H2>
<table CELLSPACING=0 CELLPADDING=3 BGCOLOR="#f5f5ff" ><tr><td>
<pre>
Standard (Mandatory) qualifiers:
[-seqall] seqall Protein sequence(s) filename and optional
format, or reference (input USA)
-menu menu [1] Select number (Values: 1 (Trypsin); 2
(Lys-C); 3 (Arg-C); 4 (Asp-N); 5
(V8-bicarb); 6 (V8-phosph); 7
(Chymotrypsin); 8 (CNBr))
[-outfile] report [*.digest] Output report file name
Additional (Optional) qualifiers: (none)
Advanced (Unprompted) qualifiers:
-aadata datafile [Eamino.dat] Molecular weight data for amino
acids
-unfavoured boolean Trypsin will not normally cut after a K if
it is followed by (e.g.) another K or a P.
Specifying this shows those cuts. as well as
the favoured ones.
-ragging boolean Allows semi-specific and non-specific
digestion. This option is particularly
useful for generating lists of peptide
sequences for protein identification using
mass-spectrometry.
-termini menu [1] Select number (Values: 1 (none); 2
(nterm); 3 (cterm); 4 (nterm OR cterm))
-overlap boolean Used for partial digestion. Shows all cuts
from favoured cut sites plus 1..3, 2..4,
3..5 etc but not (e.g.) 2..5. Overlaps are
therefore fragments with exactly one
potential cut site within it.
-allpartials boolean As for overlap but fragments containing more
than one potential cut site are included.
Associated qualifiers:
"-seqall" associated qualifiers
-sbegin1 integer Start of each sequence to be used
-send1 integer End of each sequence to be used
-sreverse1 boolean Reverse (if DNA)
-sask1 boolean Ask for begin/end/reverse
-snucleotide1 boolean Sequence is nucleotide
-sprotein1 boolean Sequence is protein
-slower1 boolean Make lower case
-supper1 boolean Make upper case
-sformat1 string Input sequence format
-sdbname1 string Database name
-sid1 string Entryname
-ufo1 string UFO features
-fformat1 string Features format
-fopenfile1 string Features file name
"-outfile" associated qualifiers
-rformat2 string Report format
-rname2 string Base file name
-rextension2 string File name extension
-rdirectory2 string Output directory
-raccshow2 boolean Show accession number in the report
-rdesshow2 boolean Show description in the report
-rscoreshow2 boolean Show the score in the report
-rusashow2 boolean Show the full USA in the report
-rmaxall2 integer Maximum total hits to report
-rmaxseq2 integer Maximum hits to report for one sequence
General qualifiers:
-auto boolean Turn off prompts
-stdout boolean Write standard output
-filter boolean Read standard input, write standard output
-options boolean Prompt for standard and additional values
-debug boolean Write debug output to program.dbg
-verbose boolean Report some/full command line options
-help boolean Report command line options. More
information on associated and general
qualifiers can be found with -help -verbose
-warning boolean Report warnings
-error boolean Report errors
-fatal boolean Report fatal errors
-die boolean Report dying program messages
</pre>
</td></tr></table>
<P>
<table border cellspacing=0 cellpadding=3 bgcolor="#ccccff">
<tr bgcolor="#FFFFCC">
<th align="left" colspan=2>Standard (Mandatory) qualifiers</th>
<th align="left">Allowed values</th>
<th align="left">Default</th>
</tr>
<tr>
<td>[-seqall]<br>(Parameter 1)</td>
<td>Protein sequence(s) filename and optional format, or reference (input USA)</td>
<td>Readable sequence(s)</td>
<td><b>Required</b></td>
</tr>
<tr>
<td>-menu</td>
<td>Select number</td>
<td><table><tr><td>1</td> <td><i>(Trypsin)</i></td></tr><tr><td>2</td> <td><i>(Lys-C)</i></td></tr><tr><td>3</td> <td><i>(Arg-C)</i></td></tr><tr><td>4</td> <td><i>(Asp-N)</i></td></tr><tr><td>5</td> <td><i>(V8-bicarb)</i></td></tr><tr><td>6</td> <td><i>(V8-phosph)</i></td></tr><tr><td>7</td> <td><i>(Chymotrypsin)</i></td></tr><tr><td>8</td> <td><i>(CNBr)</i></td></tr></table></td>
<td>1</td>
</tr>
<tr>
<td>[-outfile]<br>(Parameter 2)</td>
<td>Output report file name</td>
<td>Report output file</td>
<td><i><*></i>.digest</td>
</tr>
<tr bgcolor="#FFFFCC">
<th align="left" colspan=2>Additional (Optional) qualifiers</th>
<th align="left">Allowed values</th>
<th align="left">Default</th>
</tr>
<tr>
<td colspan=4>(none)</td>
</tr>
<tr bgcolor="#FFFFCC">
<th align="left" colspan=2>Advanced (Unprompted) qualifiers</th>
<th align="left">Allowed values</th>
<th align="left">Default</th>
</tr>
<tr>
<td>-aadata</td>
<td>Molecular weight data for amino acids</td>
<td>Data file</td>
<td>Eamino.dat</td>
</tr>
<tr>
<td>-unfavoured</td>
<td>Trypsin will not normally cut after a K if it is followed by (e.g.) another K or a P. Specifying this shows those cuts. as well as the favoured ones.</td>
<td>Boolean value Yes/No</td>
<td>No</td>
</tr>
<tr>
<td>-ragging</td>
<td>Allows semi-specific and non-specific digestion. This option is particularly useful for generating lists of peptide sequences for protein identification using mass-spectrometry.</td>
<td>Boolean value Yes/No</td>
<td>No</td>
</tr>
<tr>
<td>-termini</td>
<td>Select number</td>
<td><table><tr><td>1</td> <td><i>(none)</i></td></tr><tr><td>2</td> <td><i>(nterm)</i></td></tr><tr><td>3</td> <td><i>(cterm)</i></td></tr><tr><td>4</td> <td><i>(nterm OR cterm)</i></td></tr></table></td>
<td>1</td>
</tr>
<tr>
<td>-overlap</td>
<td>Used for partial digestion. Shows all cuts from favoured cut sites plus 1..3, 2..4, 3..5 etc but not (e.g.) 2..5. Overlaps are therefore fragments with exactly one potential cut site within it.</td>
<td>Boolean value Yes/No</td>
<td>No</td>
</tr>
<tr>
<td>-allpartials</td>
<td>As for overlap but fragments containing more than one potential cut site are included.</td>
<td>Boolean value Yes/No</td>
<td>No</td>
</tr>
</table>
<H2>
Input file format
</H2>
Any protein sequence.
<p>
<a name="input.1"></a>
<h3>Input files for usage example </h3>
'tsw:opsd_human' is a sequence entry in the example protein database 'tsw'
<p>
<p><h3>Database entry: tsw:opsd_human</h3>
<table width="90%"><tr><td bgcolor="#FFCCFF">
<pre>
ID OPSD_HUMAN Reviewed; 348 AA.
AC P08100; Q16414; Q2M249;
DT 01-AUG-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1988, sequence version 1.
DT 20-MAR-2007, entry version 91.
DE Rhodopsin (Opsin-2).
GN Name=RHO; Synonyms=OPN2;
OS Homo sapiens (Human).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;
OC Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini;
OC Catarrhini; Hominidae; Homo.
OX NCBI_TaxID=9606;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX MEDLINE=84272729; PubMed=6589631;
RA Nathans J., Hogness D.S.;
RT "Isolation and nucleotide sequence of the gene encoding human
RT rhodopsin.";
RL Proc. Natl. Acad. Sci. U.S.A. 81:4851-4855(1984).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Suwa M., Sato T., Okouchi I., Arita M., Futami K., Matsumoto S.,
RA Tsutsumi S., Aburatani H., Asai K., Akiyama Y.;
RT "Genome-wide discovery and analysis of human seven transmembrane helix
RT receptor genes.";
RL Submitted (JUL-2001) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC TISSUE=Retina;
RG The German cDNA consortium;
RL Submitted (JUN-2003) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA
RT project: the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [5]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-120.
RX PubMed=8566799; DOI=10.1016/0378-1119(95)00688-5;
RA Bennett J., Beller B., Sun D., Kariko K.;
RT "Sequence analysis of the 5.34-kb 5' flanking region of the human
RT rhodopsin-encoding gene.";
RL Gene 167:317-320(1995).
RN [6]
RP REVIEW ON RP4 VARIANTS.
RX MEDLINE=94004905; PubMed=8401533;
RA Al-Maghtheh M., Gregory C., Inglehearn C., Hardcastle A.,
RA Bhattacharya S.;
<font color=red> [Part of this file has been deleted for brevity]</font>
FT /FTId=VAR_004816.
FT VARIANT 209 209 V -> M (effect not known).
FT /FTId=VAR_004817.
FT VARIANT 211 211 H -> P (in RP4).
FT /FTId=VAR_004818.
FT VARIANT 211 211 H -> R (in RP4).
FT /FTId=VAR_004819.
FT VARIANT 216 216 M -> K (in RP4).
FT /FTId=VAR_004820.
FT VARIANT 220 220 F -> C (in RP4).
FT /FTId=VAR_004821.
FT VARIANT 222 222 C -> R (in RP4).
FT /FTId=VAR_004822.
FT VARIANT 255 255 Missing (in RP4).
FT /FTId=VAR_004823.
FT VARIANT 264 264 Missing (in RP4).
FT /FTId=VAR_004824.
FT VARIANT 267 267 P -> L (in RP4).
FT /FTId=VAR_004825.
FT VARIANT 267 267 P -> R (in RP4).
FT /FTId=VAR_004826.
FT VARIANT 292 292 A -> E (in CSNBAD1).
FT /FTId=VAR_004827.
FT VARIANT 296 296 K -> E (in RP4).
FT /FTId=VAR_004828.
FT VARIANT 297 297 S -> R (in RP4).
FT /FTId=VAR_004829.
FT VARIANT 342 342 T -> M (in RP4).
FT /FTId=VAR_004830.
FT VARIANT 345 345 V -> L (in RP4).
FT /FTId=VAR_004831.
FT VARIANT 345 345 V -> M (in RP4).
FT /FTId=VAR_004832.
FT VARIANT 347 347 P -> A (in RP4).
FT /FTId=VAR_004833.
FT VARIANT 347 347 P -> L (in RP4; common variant).
FT /FTId=VAR_004834.
FT VARIANT 347 347 P -> Q (in RP4).
FT /FTId=VAR_004835.
FT VARIANT 347 347 P -> R (in RP4).
FT /FTId=VAR_004836.
FT VARIANT 347 347 P -> S (in RP4).
FT /FTId=VAR_004837.
SQ SEQUENCE 348 AA; 38893 MW; 6F4F6FCBA34265B2 CRC64;
MNGTEGPNFY VPFSNATGVV RSPFEYPQYY LAEPWQFSML AAYMFLLIVL GFPINFLTLY
VTVQHKKLRT PLNYILLNLA VADLFMVLGG FTSTLYTSLH GYFVFGPTGC NLEGFFATLG
GEIALWSLVV LAIERYVVVC KPMSNFRFGE NHAIMGVAFT WVMALACAAP PLAGWSRYIP
EGLQCSCGID YYTLKPEVNN ESFVIYMFVV HFTIPMIIIF FCYGQLVFTV KEAAAQQQES
ATTQKAEKEV TRMVIIMVIA FLICWVPYAS VAFYIFTHQG SNFGPIFMTI PAFFAKSAAI
YNPVIYIMMN KQFRNCMLTT ICCGKNPLGD DEASATVSKT ETSQVAPA
//
</pre>
</td></tr></table><p>
<H2>
Output file format
</H2>
<p>
The output is a standard EMBOSS report file.
<p>
The results can be output in one of several styles by using the
command-line qualifier <b>-rformat xxx</b>, where 'xxx' is replaced by
the name of the required format. The available format names are: embl,
genbank, gff, pir, swiss, trace, listfile, dbmotif, diffseq, excel,
feattable, motif, regions, seqtable, simple, srs, table, tagseq
<p>
See:
<A href="http://emboss.sf.net/docs/themes/ReportFormats.html">
http://emboss.sf.net/docs/themes/ReportFormats.html</A>
for further information on report formats.
<p>
By default <b>digest</b> writes a 'seqtable' report file.
<p>
<a name="output.1"></a>
<h3>Output files for usage example </h3>
<p><h3>File: opsd_human.digest</h3>
<table width="90%"><tr><td bgcolor="#CCFFCC">
<pre>
########################################
# Program: digest
# Rundate: Sun 15 Jul 2007 12:00:00
# Commandline: digest
# -seqall tsw:opsd_human
# Report_format: seqtable
# Report_file: opsd_human.digest
########################################
#=======================================
#
# Sequence: OPSD_HUMAN from: 1 to: 348
# HitCount: 14
#
# Complete digestion with Trypsin yields 14 fragments
#
#=======================================
Start End Mol_Weight Cterm Nterm Sequence
70 135 7129.319 R Y TPLNYILLNLAVADLFMVLGGFTSTLYTSLHGYFVFGPTGCNLEGFFATLGGEIALWSLVVLAIER
178 231 6335.495 R E YIPEGLQCSCGIDYYTLKPEVNNESFVIYMFVVHFTIPMIIIFFCYGQLVFTVK
22 69 5788.873 R T SPFEYPQYYLAEPWQFSMLAAYMFLLIVLGFPINFLTLYVTVQHKKLR
253 296 5004.085 R S MVIIMVIAFLICWVPYASVAFYIFTHQGSNFGPIFMTIPAFFAK
136 177 4600.460 R Y YVVVCKPMSNFRFGENHAIMGVAFTWVMALACAAPPLAGWSR
1 21 2257.495 . S MNGTEGPNFYVPFSNATGVVR
297 311 1728.089 K Q SAAIYNPVIYIMMNK
232 245 1490.538 K A EAAAQQQESATTQK
326 339 1403.457 K T NPLGDDEASATVSK
315 325 1186.476 R N NCMLTTICCGK
340 348 902.950 K . TETSQVAPA
249 252 503.550 K M EVTR
312 314 449.504 K N QFR
246 248 346.378 K E AEK
#---------------------------------------
#---------------------------------------
</pre>
</td></tr></table><p>
<p>
The header information contains the program name, date of run, name of
the reagent used to digest the protein and the number of fragments
reported. The header will report if complete or partial digestion was
chosen.
<p>
The rest of the file consists of columns holding the following data:
<ul>
<li> The start position of the fragment
<li> The end position of the fragment
<li> The molecular weight of the fragment
<li> The residue before the cut site ('.' if start of sequence)
<li> The residue after the second cut site ('.' if end of sequence)
<li> The sequence of the fragment.
</ul>
<H2>
Data files
</H2>
None.
<H2>
Notes
</H2>
None.
<H2>
References
</H2>
None.
<H2>
Warnings
</H2>
None.
<H2>
Diagnostic Error Messages
</H2>
None.
<H2>
Exit status
</H2>
It always exits with a status of 0.
<H2>
Known bugs
</H2>
None.
<h2><a name="See also">See also</a></h2>
<table border cellpadding=4 bgcolor="#FFFFF0">
<tr><th>Program name</th><th>Description</th></tr>
<tr>
<td><a href="antigenic.html">antigenic</a></td>
<td>Finds antigenic sites in proteins</td>
</tr>
<tr>
<td><a href="epestfind.html">epestfind</a></td>
<td>Finds PEST motifs as potential proteolytic cleavage sites</td>
</tr>
<tr>
<td><a href="fuzzpro.html">fuzzpro</a></td>
<td>Protein pattern search</td>
</tr>
<tr>
<td><a href="fuzztran.html">fuzztran</a></td>
<td>Protein pattern search after translation</td>
</tr>
<tr>
<td><a href="helixturnhelix.html">helixturnhelix</a></td>
<td>Report nucleic acid binding motifs</td>
</tr>
<tr>
<td><a href="oddcomp.html">oddcomp</a></td>
<td>Find protein sequence regions with a biased composition</td>
</tr>
<tr>
<td><a href="patmatdb.html">patmatdb</a></td>
<td>Search a protein sequence with a motif</td>
</tr>
<tr>
<td><a href="patmatmotifs.html">patmatmotifs</a></td>
<td>Search a PROSITE motif database with a protein sequence</td>
</tr>
<tr>
<td><a href="pepcoil.html">pepcoil</a></td>
<td>Predicts coiled coil regions</td>
</tr>
<tr>
<td><a href="preg.html">preg</a></td>
<td>Regular expression search of a protein sequence</td>
</tr>
<tr>
<td><a href="pscan.html">pscan</a></td>
<td>Scans proteins using PRINTS</td>
</tr>
<tr>
<td><a href="sigcleave.html">sigcleave</a></td>
<td>Reports protein signal cleavage sites</td>
</tr>
</table>
<H2>
Author(s)
</H2>
Alan Bleasby (ajb © ebi.ac.uk)
<br>
European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD, UK
<H2>
History
</H2>
Written (1999) - Alan Bleasby
<H2>
Target users
</H2>
This program is intended to be used by everyone and everything, from naive users to embedded scripts.
<H2>
Comments
</H2>
None
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