The server requires protein or peptide sequence(s), depending on the selected “Analyse Mode”, in FASTA format and cannot handle nucleic acid sequences. The server is also limited to the 20 standard amino acids.
Predicts the activity directly from the peptides provide (no cutting up the sequences as with the other modes). The settings of Minimum and Maximum peptide length do not affect which peptides get predicted, but a standard limit of 2-30 residues is used for every peptide.
Each input protein is cut into every possible peptide with the given lengths (the settings of Minimum and Maximum peptide length). This exhaustive mode thereby gives a comprehensive prediction of all peptides embedded in the proteins. To make the results easier to interpret, it is noted which proteins each sequence is present in and output files also include clustering of similar sequences (70% sequence identity).
In the case of many, large proteins, this mode might give to overwhelming results and the Protein(s) w/ Proteolysis Mode is in these cases recommended.
The proteins are cleaved by combination of selected proteases (at least one must be selected). Resulting peptides outside given lengths (the settings of Minimum and Maximum peptide length) are deselected from the output. This mode considers that all cleavage sites are not always cleaved. All combinations of peptides being cleaved or not, are therefore included, to give the best overall results.
The cleavage specificities of each protease adhere to the following defined guidelines: Expasy Guidelines.