9 research outputs found
Computational approaches to protein ligand interactions: Protein haem complexes
Many different proteins bind and utilise haem to perform important biological functions; the aim of this work was to gain an understanding of some of the underlying molecular recognition processes. We considered how the protein environment determines haem binding, and what are the consequences for ligand conformation. Sets of homologous globins and of non-homologous haem-binding proteins were chosen on the basis of sequence and structural similarity, allowing us to compare ligand conformation and to explore the factors that modulate its structure. Comparison of bound and un-bound haems revealed a conformational disparity between the data-sets, suggesting that protein structural factors provide the dominant effect over the conformational features of the ligand. Even in the homologous globins, the ligand side-chain conformation is variable; greater variability within the non-homologous group reflects different local amino acid sequences of their binding pockets. Moreover, the haem skeleton can be severely distorted, being particularly sensitive to local environment, to attached molecules, and to ligation state. The haem environment was analysed to understand its role in the functional and structural variability of the ligand. Predominance of mainly-alpha structures characterises haemoproteins. While the binding sites are radically different in topology, there are preferred binding modes, with the ligand side-chains engaged in a network of hydrophobic and hydrogen bonding interactions. The stacking of aromatic side-chains on the haem skeleton also appears to be essential in the formation of protein-ligand complexes. Analysis of the haem interface revealed that specific residues prefer to line the binding site and to form ligand contacts. An attempt to correlate properties of the unrelated protein-ligand complexes with haem redox potential was made. A compendium of these analyses has been developed and made accessible via the WWW, providing a user-friendly interface for analysing protein-haem interactions. A search tool allows on-the-fly analysis of protein-ligand relationships, which should facilitate both the comparison of different binding sites, and prediction and design of novel ones
DB-PABP: a database of polyanion-binding proteins
The interactions between polyanions (PAs) and polyanion-binding proteins (PABPs) have been found to play significant roles in many essential biological processes including intracellular organization, transport and protein folding. Furthermore, many neurodegenerative disease-related proteins are PABPs. Thus, a better understanding of PA/PABP interactions may not only enhance our understandings of biological systems but also provide new clues to these deadly diseases. The literature in this field is widely scattered, suggesting the need for a comprehensive and searchable database of PABPs. The DB-PABP is a comprehensive, manually curated and searchable database of experimentally characterized PABPs. It is freely available and can be accessed online at http://pabp.bcf.ku.edu/DB_PABP/. The DB-PABP was implemented as a MySQL relational database. An interactive web interface was created using Java Server Pages (JSP). The search page of the database is organized into a main search form and a section for utilities. The main search form enables custom searches via four menus: protein names, polyanion names, the source species of the proteins and the methods used to discover the interactions. Available utilities include a commonality matrix, a function of listing PABPs by the number of interacting polyanions and a string search for author surnames. The DB-PABP is maintained at the University of Kansas. We encourage users to provide feedback and submit new data and references
Subdivision of the helix-turn-helix GntR family of bacterial regulators in the FadR, HutC, MocR, and YtrA subfamilies
Haydon and Guest (Haydon, D. J, and Guest, J. R. (1991) FEMS Microbiol Lett. 63, 291-295) first described the helix-turn-helix GntR family of bacterial regulators. They presented them as transcription factors sharing a similar N-terminal DNA-binding (D-b) domain, but they observed near-maximal divergence in the C-terminal effector-binding and oligomerization (E-b/O) domain. To elucidate this C-terminal heterogeneity, structural, phylogenetic, and functional analyses were performed on a family that now comprises about 270 members. Our comparative study first focused on the C-terminal E-b/O domains and next on DNA-binding domains and palindromic operator sequences, has classified the GntR members into four subfamilies that we called FadR, HutC, MocR, and YtrA. Among these subfamilies a degree of similarity of about 55% was observed throughout the entire sequence. Structure/function associations were highlighted although they were not absolutely stringent. The consensus sequences deduced for the DNA-binding domain were slightly different for each subfamily, suggesting that fusion between the D-b and E-b/O domains have occurred separately, with each subfamily having its own D-b domain ancestor. Moreover, the compilation of the known or predicted palindromic cis-acting elements has highlighted different operator sequences according to our subfamily subdivision. The observed C-terminal E-b/O domain heterogeneity was therefore reflected on the DNA-binding domain and on the cis-acting elements, suggesting the existence of a tight link between the three regions involved in the regulating process.Peer reviewe
A Web-based classification system of DNA-binding protein families
Rational classification of proteins encoded in sequenced genomes is
critical for making the genome sequences maximally useful for functional
and evolutionary studies. The family of DNA-binding proteins is one of
the most populated and studied amongst the various genomes of bacteria,
archaea and eukaryotes and the Web-based system presented here is an
approach to their classification. The DnaProt resource is an annotated
and searchable collection of protein sequences for the families of
DNA-binding proteins. The database contains 3238 full-length sequences
(retrieved from the SWISS-PROT database, release 38) that include, at
least, a DNA-binding domain. Sequence entries are organized into
families defined by PROSITE patterns, PRINTS motifs and de novo excised
signatures. Combining global similarities and functional motifs into a
single classification scheme, DNA-binding proteins are classified into
33 unique classes, which helps to reveal comprehensive family
relationships. To maximize family information retrieval, DnaProt
contains a collection of multiple alignments for each DNA-binding family
while the recognized motifs can be used as diagnostically functional
fingerprints. All available structural class representatives have been
referenced. The resource was developed as a Web-based management system
for online free access of customized data sets. Entries are fully
hyperlinked to facilitate easy retrieval of the original records from
the source databases while functional and phylogenetic annotation will
be applied to newly sequenced genomes. The database is freely available
for online search of a library containing specific patterns of the
identified DNA-binding protein classes and retrieval of individual
entries from our WWW server (http://kronos.biol.uoa.gr/similar to
mariak/dbDNA.html)
Protein folds and functions
Background: The recent rapid increase in the number of available three-dimensional protein structures has further highlighted the necessity to understand the relationship between biological function and structure. Using structural classification schemes such as SCOP, CATH and DALI, it is now possible to explore global relationships between protein fold and function, something which was previously impractical. Results: Using a relational database of CATH data we have generated fold distributions for arbitrary selections of proteins automatically. These distributions have been examined in the light of protein function and bound ligand. Different enzyme classes are not clearly reflected in distributions of protein class and architecture, whereas the type of bound ligand has a much more dramatic effect. Conclusions: The availability of structural classification data has enabled this novel overview analysis. We conclude that function at the top level of the EC number enzyme classification is not related to fold, as only a very few specific residues are actually responsible for enzyme activity. Conversely, the fold is much more closely related to ligand type
The genome sequence of Bifidobacterium longum reflects its adaptation to the human gastrointestinal tract
Bifidobacteria are Gram-positive prokaryotes that naturally colonize the human gastrointestinal tract (GIT) and vagina. Although not numerically dominant in the complex intestinal microflora, they are considered as key commensals that promote a healthy GIT. We determined the 2.26-Mb genome sequence of an infant-derived strain of Bifidobacterium longum, and identified 1,730 possible coding sequences organized in a 60%–GC circular chromosome. Bioinformatic analysis revealed several physiological traits that could partially explain the successful adaptation of this bacteria to the colon. An unexpectedly large number of the predicted proteins appeared to be specialized for catabolism of a variety of oligosaccharides, some possibly released by rare or novel glycosyl hydrolases acting on “nondigestible” plant polymers or host-derived glycoproteins and glycoconjugates. This ability to scavenge from a large variety of nutrients likely contributes to the competitiveness and persistence of bifidobacteria in the colon. Many genes for oligosaccharide metabolism were found in self-regulated modules that appear to have arisen in part from gene duplication or horizontal acquisition. Complete pathways for all amino acids, nucleotides, and some key vitamins were identified; however, routes for Asp and Cys were atypical. More importantly, genome analysis provided insights into the reciprocal interactions of bifidobacteria with their hosts. We identified polypeptides that showed homology to most major proteins needed for production of glycoprotein-binding fimbriae, structures that could possibly be important for adhesion and persistence in the GIT. We also found a eukaryotic-type serine protease inhibitor (serpin) possibly involved in the reported immunomodulatory activity of bifidobacteria