W284–W288 Nucleic Acids Research, 2005, Vol. 33, Web Server issue doi:10.1093/nar/gki418
FFAS03: a server for profile–profile sequence alignments Lukasz Jaroszewski, Leszek Rychlewski1, Zhanwen Li, Weizhong Li and Adam Godzik* Bioinformatics Program, The Burnham Institute, 10901 N. Torrey Pines Road, La Jolla, CA 92037, USA and 1 BioInfoBank Institute, ul. Limanowskiego 24 A, 60-744 Poznan, Poland Received February 17, 2005; Revised and Accepted March 21, 2005
ABSTRACT The FFAS03 server provides a web interface to the third generation of the profile–profile alignment and fold-recognition algorithm of fold and function assignment system (FFAS) [L. Rychlewski, L. Jaroszewski, W. Li and A. Godzik (2000), Protein Sci., 9, 232–241]. Profile–profile algorithms use information present in sequences of homologous proteins to amplify the patterns defining the family. As a result, they enable detection of remote homologies beyond the reach of other methods. FFAS, initially developed in 2000, is consistently one of the best ranked fold prediction methods in the CAFASP and LiveBench competitions. It is also used by several fold-recognition consensus methods and meta-servers. The FFAS03 server accepts a user supplied protein sequence and automatically generates a profile, which is then compared with several sets of sequence profiles of proteins from PDB, COG, PFAM and SCOP. The profile databases used by the server are automatically updated with the latest structural and sequence information. The server provides access to the alignment analysis, multiple alignment, and comparative modeling tools. Access to the server is open for both academic and commercial researchers. The FFAS03 server is available at http://ffas.burnham.org.
INTRODUCTION The most effective methods of protein structure and function predictions are based on establishing a homology between the protein of interest and an already characterized protein. The standard sequence–sequence comparison methods, however, rapidly lose sensitivity in the ‘twilight zone’ of 30% or less
sequence identity (1). The sensitivity of homology recognition can be improved by using information present in the families of protein sequences connected with detectable homology. In this approach, one compares a protein sequence with a protein family represented by a sequence profile [e.g. in PSI-BLAST (2)]. A next step in this strategy is to compare two sequence profiles. The fold and function assignment system (FFAS) is a profile–profile comparison algorithm developed in 2000 by our group (3). Profile–profile scoring was used earlier to align short blocks (4), and FFAS extended this approach to allow for gaps and align entire proteins. Profile–profile alignment algorithms surpass sequence-sequence and profile-sequence alignment algorithms in terms of sensitivity (3) and alignment accuracy (5). FFAS is regularly assessed in CASP (6) and CAFASP (7) competitions and continually benchmarked in LiveBench (8) experiment. In the last LiveBench, it was ranked as the most sensitive of all sequence-based methods in the category of difficult fold prediction (see http://bioinfo.pl/ Meta/results.pl?B=LiveBench&V=9). Development of FFAS was followed by many similar methods that differ in the way two profiles are compared with each other (9–13). FFAS ALGORITHM Each profile–profile alignment method includes four steps: (i) preparation of the multiple sequence alignment, (ii) calculation of a profile, (iii) alignment of profile with sequence profiles from the database such as PDB and (iv) estimation of the statistical significance of the alignment score. In FFAS method, the multiple sequence alignment is prepared using PSI-BLAST (2). Five iterations of PSI-BLAST are performed against the NR85S database of protein sequences (NR85S database is described in Table 1). In the second step, all sequences found by PSI-BLAST with E-value < 0.005 are used for profile calculation. Weights are assigned to sequences based on their similarity to other sequences in the multiple sequence alignment (3).
*To whom correspondence should be addressed. Tel: +1 858 646 3168; Fax: +1 858 713 9925; Email:
[email protected] Present address: Lukasz Jaroszewski, Joint Center for Structural Genomics, UCSD, La Jolla, CA 92093, USA ª The Author 2005. Published by Oxford University Press. All rights reserved. The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact
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Nucleic Acids Research, 2005, Vol. 33, Web Server issue
Table 1. The databases used by the FFAS03 server Database
Source of data
Preparation
NR85S NCBI, SEED (sequences)
PDB (profiles) PFAM (profiles) COG (profiles) SCOP (profiles)
JCSG (profiles)
Protein sequences from the NCBI NR database and predicted open reading frames from unfinished bacterial genomes (kindly provided by Ross Overbeek) are clustered at 85% of sequence identity with the CD-HIT program (15). Regions of low complexity are masked with SEG (16). Protein Data FFAS profiles of all unique proteins Bank (clustered at 99% identity level) from the PDB (17), including prereleased entries. PFAM website FFAS profiles of all PFAM (18) domains longer than 25 residues. NCBI FFAS profiles of all domains from COG database longer than 25 residues (19). SCOP–ASTRAL FFAS profiles of SCOP domain website sequences with
