Protein to protein sequence comparisons
are more sensitive than DNA to protein sequence comparisons. After
using the 6 Frame Translation program,
"cut" out the correct translation and "paste"
it in the appropriate search window (see below).
WU-BLASTP+BEAUTY / nr
protein - Warren Gish's BLAST with gapped alignments with
BEAUTY post-processing (WU/BCM). [H]
[O]
[P]
[E]
Make sure the selection circle/diamond
is clicked.
Enter your amino acid sequence in the input box.
Click "Perform search".
Notes on this type of search:
WU-BLASTX
(Washington University BLASTX)is a search program that compares
a DNA sequence, translated in all possible reading frames,
to a selected protein database. Thus, it compares six possible
translation products of a nucleotide query sequence (three translations,
both strands). Translating the DNA sequence prior to searching
greatly improves the accuracy of the search, as true sequence
similarity is much easier to detect in proteins that in DNA. In
DNA, the rapidly mutating third base position of each codon and
the large non-coding regions limit the effectiveness of direct
DNA sequence comparisons in detecting non-identical matches.
Other programs that search translated
DNA queries against protein databases include BLASTX
2.0 from NCBI and FASTX
(University of Virginina).
BLAST uses
a heuristicsearch algorithm, starting with a quick, but
less accurate, exact-match procedure to identify possible matches
from the entire database, and then using the more sensitive but
time-consuming Smith-Waterman algorithm to search these matches
and create the final output. Details
of BLAST search strategy
"with gapped alignments"
means that this search engine
recognizes gaps in sequence alignments and incorporates gap
costs cooresponding to these areas in the final sequence alignment
score. Generally, the gap cost charges a large initial penality
for the existence of a gap, and smaller penalities for each individual
residue. This takes into account that each mutational event can
insert or delete multiple residues at a time, and thus the bulk
of the gap cost penalty is for the existence of the mutation itself,
not the length. This also allows matches to extend across short
gaps and still appear as contiguous in the sequence alignment.
BEAUTY (BLAST Enhanced Alignment Utility)
Post- Processor incorporates additional
information directly into the BLASTX search output, including
information on the locations of any annotated domains among the
sequence matches produced by the BLASTX search. Regions of sequence
matches (local hits) that overlap annotated domains may have the
same function as the domain. Thus, this information improves the
identification of weak, but functional significant sequence matches.
Example
of BEAUTY output
filters:This search uses XNU+SEG filters for all searches (click on [P]
in the search line for this information.) These filters convert
low-complexity protein regions (such as runs of Q's) to X's --
see the description of the '-filter' option in the BLAST
Help Pages.