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digest |
Function
Protein proteolytic enzyme or reagent cleavage digestDescription
digest finds the positions where a specified proteolytic enzyme or reagent might cut a peptide sequence.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.
Trypsin will not normally cut after a K if it is followed by (e.g.) another K or a P. Specifying the qualifier -unfavoured shows those cuts. as well as the favoured ones.
If you wish to emulate a partial digestion, then using the -overlap 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.
i.e. fragments containing one potential cut site are also shown.
If you wish to emulate a very partial digestion (!) then using the -allpartials qualifier will do what -overlap 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.
i.e. fragments containing one or more potential cut site are also shown.
Usage
Here is a sample session with digest
% digest
Protein proteolytic enzyme or reagent cleavage digest
Input sequence: tsw:opsd_human
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]:
Output report [opsd_human.digest]:
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Go to the input files for this example
Go to the output files for this example
Command line arguments
Standard (Mandatory) qualifiers:
[-sequence] sequence Sequence USA
-menu menu Select number
[-outfile] report Output report file name
Additional (Optional) qualifiers: (none)
Advanced (Unprompted) qualifiers:
-aadata datafile 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.
-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:
"-sequence" associated qualifiers
-sbegin1 integer Start of the sequence to be used
-send1 integer End of the 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
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 deaths
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| Standard (Mandatory) qualifiers | Allowed values | Default | |||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| [-sequence] (Parameter 1) |
Sequence USA | Readable sequence | Required | ||||||||||||||||
| -menu | Select number |
|
1 | ||||||||||||||||
| [-outfile] (Parameter 2) |
Output report file name | Report output file | |||||||||||||||||
| Additional (Optional) qualifiers | Allowed values | Default | |||||||||||||||||
| (none) | |||||||||||||||||||
| Advanced (Unprompted) qualifiers | Allowed values | Default | |||||||||||||||||
| -aadata | Molecular weight data for amino acids | Data file | Eamino.dat | ||||||||||||||||
| -unfavoured | 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. | Boolean value Yes/No | No | ||||||||||||||||
| -overlap | 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. | Boolean value Yes/No | No | ||||||||||||||||
| -allpartials | As for overlap but fragments containing more than one potential cut site are included. | Boolean value Yes/No | No | ||||||||||||||||
Input file format
Any protein sequence.Input files for usage example
'tsw:opsd_human' is a sequence entry in the example protein database 'tsw'
Database entry: tsw:opsd_human
ID OPSD_HUMAN STANDARD; PRT; 348 AA.
AC P08100; Q16414;
DT 01-AUG-1988 (Rel. 08, Created)
DT 01-AUG-1988 (Rel. 08, Last sequence update)
DT 15-JUL-1999 (Rel. 38, Last annotation update)
DE RHODOPSIN.
GN RHO.
OS Homo sapiens (Human).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Mammalia;
OC Eutheria; Primates; Catarrhini; Hominidae; Homo.
RN [1]
RP SEQUENCE FROM N.A.
RX MEDLINE; 84272729.
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 SEQUENCE OF 1-120 FROM N.A.
RA BENNETT J., BELLER B., SUN D., KARIKO K.;
RL Submitted (NOV-1994) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP REVIEW ON ADRP VARIANTS.
RX MEDLINE; 94004905.
RA AL-MAGHTHEH M., GREGORY C., INGLEHEARN C., HARDCASTLE A.,
RA BHATTACHARYA S.;
RT "Rhodopsin mutations in autosomal dominant retinitis pigmentosa.";
RL Hum. Mutat. 2:249-255(1993).
RN [4]
RP VARIANT ADRP HIS-23.
RX MEDLINE; 90136922.
RA DRYJA T.P., MCGEE T.L., REICHEI E., HAHN L.B., COWLEY G.S.,
RA YANDELL D.W., SANDBERG M.A., BERSON E.L.;
RT "A point mutation of the rhodopsin gene in one form of retinitis
RT pigmentosa.";
RL Nature 343:364-366(1990).
RN [5]
RP VARIANTS ADRP.
RX MEDLINE; 91051574.
RA FARRAR G.J., KENNA P., REDMOND R., MCWILLIAM P., BRADLEY D.G.,
RA HUMPHRIES M.M., SHARP E.M., INGLEHEARN C.F., BASHIR R., JAY M.,
RA WATTY A., LUDWIG M., SCHINZEL A., SAMANNS C., GAL A.,
RA BHATTACHARYA S.S., HUMPHRIES P.;
RT "Autosomal dominant retinitis pigmentosa: absence of the rhodopsin
RT proline-->histidine substitution (codon 23) in pedigrees from
RT Europe.";
RL Am. J. Hum. Genet. 47:941-945(1990).
RN [6]
RP VARIANTS ADRP HIS-23; ARG-58; LEU-347 AND SER-347.
RX MEDLINE; 91015273.
[Part of this file has been deleted for brevity]
FT /FTId=VAR_004816.
FT VARIANT 209 209 V -> M (EFFECT NOT KNOWN).
FT /FTId=VAR_004817.
FT VARIANT 211 211 H -> P (IN ADRP).
FT /FTId=VAR_004818.
FT VARIANT 211 211 H -> R (IN ADRP).
FT /FTId=VAR_004819.
FT VARIANT 216 216 M -> K (IN ADRP).
FT /FTId=VAR_004820.
FT VARIANT 220 220 F -> C (IN ADRP).
FT /FTId=VAR_004821.
FT VARIANT 222 222 C -> R (IN ADRP).
FT /FTId=VAR_004822.
FT VARIANT 255 255 MISSING (IN ADRP).
FT /FTId=VAR_004823.
FT VARIANT 264 264 MISSING (IN ADRP).
FT /FTId=VAR_004824.
FT VARIANT 267 267 P -> L (IN ADRP).
FT /FTId=VAR_004825.
FT VARIANT 267 267 P -> R (IN ADRP).
FT /FTId=VAR_004826.
FT VARIANT 292 292 A -> E (IN CSNB4).
FT /FTId=VAR_004827.
FT VARIANT 296 296 K -> E (IN ADRP).
FT /FTId=VAR_004828.
FT VARIANT 297 297 S -> R (IN ADRP).
FT /FTId=VAR_004829.
FT VARIANT 342 342 T -> M (IN ADRP).
FT /FTId=VAR_004830.
FT VARIANT 345 345 V -> L (IN ADRP).
FT /FTId=VAR_004831.
FT VARIANT 345 345 V -> M (IN ADRP).
FT /FTId=VAR_004832.
FT VARIANT 347 347 P -> A (IN ADRP).
FT /FTId=VAR_004833.
FT VARIANT 347 347 P -> L (IN ADRP; COMMON VARIANT).
FT /FTId=VAR_004834.
FT VARIANT 347 347 P -> Q (IN ADRP).
FT /FTId=VAR_004835.
FT VARIANT 347 347 P -> R (IN ADRP).
FT /FTId=VAR_004836.
FT VARIANT 347 347 P -> S (IN ADRP).
FT /FTId=VAR_004837.
SQ SEQUENCE 348 AA; 38892 MW; 07443BEA CRC32;
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
//
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Output file format
The output is a standard EMBOSS report file.
The results can be output in one of several styles by using the command-line qualifier -rformat xxx, 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
See: http://emboss.sf.net/docs/themes/ReportFormats.html for further information on report formats.
By default digest writes a 'seqtable' report file.
Output files for usage example
File: opsd_human.digest
########################################
# Program: digest
# Rundate: Fri Jul 15 2005 12:00:00
# 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
#---------------------------------------
#---------------------------------------
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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.
The rest of the file consists of columns holding the following data:
- The start position of the fragment
- The end position of the fragment
- The molecular weight of the fragment
- The residue before the cut site ('.' if start of sequence)
- The residue after the second cut site ('.' if end of sequence)
- The sequence of the fragment.
Data files
None.Notes
None.References
None.Warnings
None.Diagnostic Error Messages
None.Exit status
It always exits with a status of 0.Known bugs
None.See also
| Program name | Description |
|---|---|
| antigenic | Finds antigenic sites in proteins |
| epestfind | Finds PEST motifs as potential proteolytic cleavage sites |
| fuzzpro | Protein pattern search |
| fuzztran | Protein pattern search after translation |
| helixturnhelix | Report nucleic acid binding motifs |
| oddcomp | Find protein sequence regions with a biased composition |
| patmatdb | Search a protein sequence with a motif |
| patmatmotifs | Search a PROSITE motif database with a protein sequence |
| pepcoil | Predicts coiled coil regions |
| preg | Regular expression search of a protein sequence |
| pscan | Scans proteins using PRINTS |
| sigcleave | Reports protein signal cleavage sites |
Author(s)
Alan Bleasby (ajb © ebi.ac.uk)European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD, UK