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A simple approach for the sensitive detection of distant relationships among protein families and for sequence--structure alignment via comparison of hidden Markov models based on their quasi-consensus sequences is presented. Using a previously published benchmark dataset, the approach is demonstrated to give better homology detection and yield alignments with improved accuracy in comparison to an existing state-of-the-art dynamic programming profile--profile comparison method. This method also runs significantly faster and is therefore suitable for a server covering the rapidly increasing structure database. A server based on this method is available at http://liao.cis.udel.edu/website/servers/modmod Contact:roland.dunbrack@fccc.edu; lliao@mail.eecis.udel.edu

The rapid growth of protein sequence databases has necessitated the development of methods to computationally derive annotation for uncharacterized entries. Most such methods focus on "global" annotation, such as molecular function or biological process. Methods to supply high-accuracy "local" annotation to functional sites based on structural information at the level of individual amino acids are relatively rare. In this chapter we will describe a method we have developed for annotation of functional residues within experimentally-uncharacterized proteins that relies on position-specific site annotation rules (PIR Site Rules) derived from structural and experimental information. These PIR Site Rules are manually defined to allow for conditional propagation of annotation. Each rule specifies a tripartite set of conditions whereby candidates for annotation must pass a whole-protein classification test (that is, have end-to-end match to a whole-protein-based HMM), match a site-specific profile HMM and, finally, match functionally and structurally characterized residues of a template. Positive matches trigger the appropriate annotation for active site residues, binding site residues, modified residues, or other functionally important amino acids. The strict criteria used in this process have rendered high-confidence annotation suitable for UniProtKB/Swiss-Prot features.

Most computational methods for transmembrane protein topology prediction rely on compositional bias of amino acids to locate those hydrophobic domains in transmembrane proteins. Because signal peptides also contain hydrophobic segments, these computational prediction methods often misidentify signal peptides as transmembrane proteins. Here, we present a new approach that combines the SVM-Fisher discrimination method and TMMOD - a hidden Markov model based predictor for transmembrane proteins. While TMMOD alone has already outperformed most existing methods in both identification and topology prediction, this new approach further improves the ability of TMMOD to discriminate between transmembrane proteins and signal peptide containing proteins, reducing mis-prediction of signal peptides by more than 30% in our test data.

Summary: A public server for evaluating the accuracy of protein sequence alignment methods is presented. CASA is an implementation of the alignment accuracy benchmark presented by Sauder et al. (Proteins, 40, 6–22, 2000). The benchmark currently contains 39321 pairwise protein structure alignments produced with the CE program from SCOP domain definitions. The server produces graphical and tabular comparisons of the accuracy of a user’s input sequence alignments with other commonly used programs, such as BLAST, PSI-BLAST, Clustal W, and SAM-T99. Availability: The server is located at http://capb.dbi.udel.edu/casa. Contact: RL_ Dunbrack@fccc.edu * To whom correspondence should be addressed.