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Bioinformatical analysis of G-protein-coupled receptors.

G-protein-coupled receptors play a key role in cellular signaling networks that regulate various physiological processes, such as vision, smell, taste, neurotransmission, secretion, inflammatory, immune responses, cellular metabolism, and cellular growth. These proteins are very important for understanding human physiology and disease. Many efforts in pharmaceutical research have been aimed at understanding their structure and function. Unfortunately, because they are difficult to crystallize and most of them will not dissolve in normal solvents, so far very few G-protein-coupled receptor structures have been determined. In contrast, more than 1000 G-protein-coupled receptor sequences are known, and many more are expected to become known soon. In view of the extremely unbalanced state, it would be very useful to develop a fast sequence-based method to identify their different types. This would no doubt have practical value for both basic research and drug discovery because the function or binding specificity of a G-protein coupled receptor is determined by the particular type it belongs to. To realize this, a statistical analysis has been performed for 566 G-protein-coupled receptors classified into seven different types. The results indicate that the types of G-protein-coupled receptors are predictable to a considerable accurate extent if a good training data set can be established for such a goal.

Algorithms↗

Bioinformatic comparison of structures and homology-models of matrix metalloproteinases.

The entire family of human matrix metalloproteinases (MMPs) was investigated using phylogenetic trees and homology modeling. The phylogenetic analysis indicates that individual domains of each MMP have evolved in a correlated manner. Despite their high sequence similarity, the phylogenetic tree of the catalytic domains already allows functional (e.g., linked to regulation and substrate recognition) homologies between different MMPs to be identified. The same pattern of functional homologies is confirmed by the phylogenetic analysis of the mature proteins. Structural models were built for the catalytic domains of the entire MMP family, for twelve hemopexin domains and for twelve mature proteins. The surface properties around the active site cleft of the modeled and experimental structures are quite conserved, whereas the hemopexin domains are more differentiated, possibly indicating a role in determining substrate specificity. The analysis of mature MMPs showed that the area of the interface between the catalytic and hemopexin domains is essentially conserved, with both hydrophobic and hydrophilic amino acids at the interface. The absence of specific conserved interdomain contacts suggests that the interface is tolerant to amino acid replacements, and that there may be a certain degree of plasticity with respect to the reciprocal orientation of the two domains.

Binding Sites↗

Clinical and pharmacogenomic data mining: 3. Zeta theory as a general tactic for clinical bioinformatics.

A new approach, a Zeta Theory of observations, data, and data mining, is being forged from a theory of expected information into an even more cohesive and comprehensive form by the challenge of general genomic, pharmacogenomic, and proteomic data. In this paper, the focus is not on studies using the specific tool FANO (CliniMiner) but on extensions to a new broader theoretical approach, aspects of which can easily be implemented into, or otherwise support, excellent existing methods, such as forms of multivariate analysis and IBM's product Intelligent Miner. The theory should perhaps be distinguished from an existing purely number-theoretic area sometimes also known as Zeta Theory, which focuses on the Riemann Zeta Function and the ways in which it governs the distribution of prime numbers. However, Zeta Theory as used here overlaps heavily with it and actually makes use of these same matters. The distinction is that it enters from a Bayesian information theory and data representation perspective. It could thus be considered an application of the 'mathematician's version'. The application is by no means confined to areas of modern biomedicine, and indeed its generality, even merging into quantum mechanics, is a key feature. Other areas with some similar challenges as modern biology, and which have inspired data mining methods such as IBM's Intelligent Miner, include commerce. But for several reasons discussed, modern molecular biology and medicine seem particularly challenging, and this relates to the often irreducible high dimensionality of the data. This thus remains our main target.

Computational Biology↗

Mass spectrometry-based proteomics combined with bioinformatic tools for bacterial classification.

Timely classification and identification of bacteria is of vital importance in many areas of public health. We present a mass spectrometry (MS)-based proteomics approach for bacterial classification. In this method, a bacterial proteome database is derived from all potential protein coding open reading frames (ORFs) found in 170 fully sequenced bacterial genomes. Amino acid sequences of tryptic peptides obtained by LC-ESI MS/MS analysis of the digest of bacterial cell extracts are assigned to individual bacterial proteomes in the database. Phylogenetic profiles of these peptides are used to create a matrix of sequence-to-bacterium assignments. These matrixes, viewed as specific assignment bitmaps, are analyzed using statistical tools to reveal the relatedness between a test bacterial sample and the microorganism database. It is shown that, if a sufficient amount of sequence information is obtained from the MS/MS experiments, a bacterial sample can be classified to a strain level by using this proteomics method, leading to its positive identification.

Amino Acid Sequence↗

Bridging neuropeptidomics and genomics with bioinformatics: Prediction of mammalian neuropeptide prohormone processing.

Neuropeptides are an important class of cell to cell signaling molecules that are difficult to predict from genetic information because of their large number of post-translational modifications. The transition from prohormone genetic sequence information to the determination of the biologically active neuropeptides requires the identification of the cleaved basic sites, among the many possible cleavage sites, that exist in the prohormone. We report a binary logistic regression model trained on mammalian prohormones that is more sensitive than existing methods in predicting these processing sites, and demonstrate the application of this method to mammalian neuropeptidomic studies. By comparing the predictive abilities of a binary logistic model trained on molluscan prohormone cleavages with the reported model, we establish the need for phyla-specific models.

Algorithms↗

Proteomic and bioinformatic characterization of the biogenesis and function of melanosomes.

Melanin, which is responsible for virtually all visible skin, hair, and eye pigmentation in humans, is synthesized, deposited, and distributed in subcellular organelles termed melanosomes. A comprehensive determination of the protein composition of this organelle has been obstructed by the melanin present. Here, we report a novel method of removing melanin that includes in-solution digestion and immobilized metal affinity chromatography (IMAC). Together with in-gel digestion, this method has allowed us to characterize melanosome proteomes at various developmental stages by tandem mass spectrometry. Comparative profiling and functional characterization of the melanosome proteomes identified approximately 1500 proteins in melanosomes of all stages, with approximately 600 in any given stage. These proteins include 16 homologous to mouse coat color genes and many associated with human pigmentary diseases. Approximately 100 proteins shared by melanosomes from pigmented and nonpigmented melanocytes define the essential melanosome proteome. Proteins validated by confirming their intracellular localization include PEDF (pigment-epithelium derived factor) and SLC24A5 (sodium/potassium/calcium exchanger 5, NCKX5). The sharing of proteins between melanosomes and other lysosome-related organelles suggests a common evolutionary origin. This work represents a model for the study of the biogenesis of lysosome-related organelles.

Animals↗

[New genes on human chromosome 7: bioinformatic analysis of a gene cluster from the POLR2J family].

Four independent genes encoding various variants of the hRPB11 subunit of Homo sapiens RNA polymerase II were revealed in human chromosome 7. Three genes (POLR2J1, POLR2J2, and POLR2J3) form a cluster of total length 214530 bp in the genetic locus 7q22.1 on the long arm of chromosome 7 (contig NT_007933). The fourth gene (POLR2J4, 31040 bp) was localized in the cytogenetic locus 7p13 of the short arm of chromosome 7 (contig NT_007819). An analysis enabled us to refine dissimilar experimental data on the mapping of the hRPB11 subunit gene on chromosome 7. In particular, the presence of three sites of its localization according to data on hybridization with fluorescent-labeled probes (the FISH method) was explained. It was established that, upon the expression of the four human POLR2J genes, at least 14 types of mature mRNAs encoding somewhat differing hRPB11 isoforms can be synthesized. Eleven of these mRNAs were revealed (as full-length copies or clearly identifiable fragments) in the available databases of expressed sequence tags and cDNAs. The most probable scheme of origination of the multiple genes of the POLR2J family, as a result of three consecutive segmented duplications increasing in size, was proposed and substantiated. On the basis of the scheme, some assumptions on the pathways of evolution of separate human genes and the mechanisms of generation of protein diversity in higher eukaryotes were made. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 6; see also http://www.maik.ru.

Chromosome Mapping↗

Bioinformatics.

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Computational Biology↗