Clustering of protein families into functional subtypes using Relative Complexity Measure with reduced amino acid alphabets

Background: Phylogenetic analysis can be used to divide a protein family into subfamilies in the absence of experimental information. Most phylogenetic analysis methods utilize multiple alignment of sequences and are based on an evolutionary model. However, multiple alignment is not an automated procedure and requires human intervention to maintain alignment integrity and to produce phylogenies consistent with the functional splits in underlying sequences. To address this problem, we propose to use the alignment-free Relative Complexity Measure (RCM) combined with reduced amino acid alphabets to cluster protein families into functional subtypes purely on sequence criteria. Comparison with an alignment-based approach was also carried out to test the quality of the clustering. Results: We demonstrate the robustness of RCM with reduced alphabets in clustering of protein sequences into families in a simulated dataset and seven well-characterized protein datasets. On protein datasets, crotonases, mandelate racemases, nucleotidyl cyclases and glycoside hydrolase family 2 were clustered into subfamilies with 100% accuracy whereas acyl transferase domains, haloacid dehalogenases, and vicinal oxygen chelates could be assigned to subfamilies with 97.2%, 96.9% and 92.2% accuracies, respectively. Conclusions: The overall combination of methods in this paper is useful for clustering protein families into subtypes based on solely protein sequence information. The method is also flexible and computationally fast because it does not require multiple alignment of sequences

Classification of proteins using sequential and structural features

Classification of proteins is an important process in many areas of bioinformatics research. In this thesis, we devised three different strategies to classify proteins with high accuracy that may have implications for function and attribute annotation. First, protein families were classified into different functional subtypes using a classification-via-clustering approach by using relative complexity measure with reduced amino acid alphabets (RAAA). The devised procedure does not require multiple alignment of sequences and produce high classification accuracies. Second, different fixed-length motif and RAAA combinations were used as features to represent proteins from different thermostability classes. A T-test based dimensionality reduction scheme was applied to reduce the number of features and those features were used to develop support vector machine classifiers. The devised procedure produced better results with less number of features than purely using native protein alphabet. Third, a non-homologous protein structure dataset containing hyperthermophilic, thermophilic, and mesophilic proteins was assembled de novo. Comprehensive statistical analyses of the dataset were carried out to highlight novel features correlated with increased thermostability and machine learning approaches were used to discriminate the proteins. For the first time, our results strongly indicate that combined sequential and structural features are better predictors of protein thermostability than purely sequential or structural features. Furthermore, the discrimination capability of machine learning models strongly depends on RAAAs

The extracellular N-terminal domain suffices to discriminate class C G Protein-Coupled Receptor subtypes from n-grams of their sequences

The investigation of protein functionality often relies on the knowledge of crystal 3-D structure. This structure is not always known or easily unravelled, which is the case of eukaryotic cell membrane proteins such as G Protein-Coupled Receptors (GPCRs) and specially of those of class C, which are the target of the current study. In the absence of information about tertiary or quaternary structures, functionality can be investigated from the primary structure, that is, from the amino acid sequence. In previous research, we found that the different subtypes of class C GPCRs could be discriminated with a high level of accuracy from the n-gram transformation of their complete primary sequences, using a method that combined two-stage feature selection with kernel classifiers. This study aims at discovering whether subunits of the complete sequence retain such discrimination capabilities. We report experiments that show that the extracellular N-terminal domain of the receptor suffices to retain the classification accuracy of the complete sequence and that it does so using a reduced selection of n-grams whose length of up to five amino acids opens up an avenue for class C GPCR signature motif discovery.Peer ReviewedPostprint (author's final draft

Testing robustness of relative complexity measure method constructing robust phylogenetic trees for Galanthus L. Using the relative complexity measure

Background: Most phylogeny analysis methods based on molecular sequences use multiple alignment where the quality of the alignment, which is dependent on the alignment parameters, determines the accuracy of the resulting trees. Different parameter combinations chosen for the multiple alignment may result in different phylogenies. A new non-alignment based approach, Relative Complexity Measure (RCM), has been introduced to tackle this problem and proven to work in fungi and mitochondrial DNA. Result: In this work, we present an application of the RCM method to reconstruct robust phylogenetic trees using sequence data for genus Galanthus obtained from different regions in Turkey. Phylogenies have been analyzed using nuclear and chloroplast DNA sequences. Results showed that, the tree obtained from nuclear ribosomal RNA gene sequences was more robust, while the tree obtained from the chloroplast DNA showed a higher degree of variation. Conclusions: Phylogenies generated by Relative Complexity Measure were found to be robust and results of RCM were more reliable than the compared techniques. Particularly, to overcome MSA-based problems, RCM seems to be a reasonable way and a good alternative to MSA-based phylogenetic analysis. We believe our method will become a mainstream phylogeny construction method especially for the highly variable sequence families where the accuracy of the MSA heavily depends on the alignment parameters

A computational intelligence analysis of G proteincoupled receptor sequinces for pharmacoproteomic applications

Arguably, drug research has contributed more to the progress of medicine during the past decades than any other scientific factor. One of the main areas of drug research is related to the analysis of proteins. The world of pharmacology is becoming increasingly dependent on the advances in the fields of genomics and proteomics. This dependency brings about the challenge of finding robust methods to analyze the complex data they generate. Such challenge invites us to go one step further than traditional statistics and resort to approaches under the conceptual umbrella of artificial intelligence, including machine learning (ML), statistical pattern recognition and soft computing methods. Sound statistical principles are essential to trust the evidence base built through the use of such approaches. Statistical ML methods are thus at the core of the current thesis. More than 50% of drugs currently available target only four key protein families, from which almost a 30% correspond to the G Protein-Coupled Receptors (GPCR) superfamily. This superfamily regulates the function of most cells in living organisms and is at the centre of the investigations reported in the current thesis. No much is known about the 3D structure of these proteins. Fortunately, plenty of information regarding their amino acid sequences is readily available. The automatic grouping and classification of GPCRs into families and these into subtypes based on sequence analysis may significantly contribute to ascertain the pharmaceutically relevant properties of this protein superfamily. There is no biologically-relevant manner of representing the symbolic sequences describing proteins using real-valued vectors. This does not preclude the possibility of analyzing them using principled methods. These may come, amongst others, from the field of statisticalML. Particularly, kernel methods can be used to this purpose. Moreover, the visualization of high-dimensional protein sequence data can be a key exploratory tool for finding meaningful information that might be obscured by their intrinsic complexity. That is why the objective of the research described in this thesis is twofold: first, the design of adequate visualization-oriented artificial intelligence-based methods for the analysis of GPCR sequential data, and second, the application of the developed methods in relevant pharmacoproteomic problems such as GPCR subtyping and protein alignment-free analysis.Se podría decir que la investigación farmacológica ha desempeñado un papel predominante en el avance de la medicina a lo largo de las últimas décadas. Una de las áreas principales de investigación farmacológica es la relacionada con el estudio de proteínas. La farmacología depende cada vez más de los avances en genómica y proteómica, lo que conlleva el reto de diseñar métodos robustos para el análisis de los datos complejos que generan. Tal reto nos incita a ir más allá de la estadística tradicional para recurrir a enfoques dentro del campo de la inteligencia artificial, incluyendo el aprendizaje automático y el reconocimiento de patrones estadístico, entre otros. El uso de principios sólidos de teoría estadística es esencial para confiar en la base de evidencia obtenida mediante estos enfoques. Los métodos de aprendizaje automático estadístico son uno de los fundamentos de esta tesis. Más del 50% de los fármacos en uso hoy en día tienen como ¿diana¿ apenas cuatro familias clave de proteínas, de las que un 30% corresponden a la super-familia de los G-Protein Coupled Receptors (GPCR). Los GPCR regulan la funcionalidad de la mayoría de las células y son el objetivo central de la tesis. Se desconoce la estructura 3D de la mayoría de estas proteínas, pero, en cambio, hay mucha información disponible de sus secuencias de amino ácidos. El agrupamiento y clasificación automáticos de los GPCR en familias, y de éstas a su vez en subtipos, en base a sus secuencias, pueden contribuir de forma significativa a dilucidar aquellas de sus propiedades de interés farmacológico. No hay forma biológicamente relevante de representar las secuencias simbólicas de las proteínas mediante vectores reales. Esto no impide que se puedan analizar con métodos adecuados. Entre estos se cuentan las técnicas provenientes del aprendizaje automático estadístico y, en particular, los métodos kernel. Por otro lado, la visualización de secuencias de proteínas de alta dimensionalidad puede ser una herramienta clave para la exploración y análisis de las mismas. Es por ello que el objetivo central de la investigación descrita en esta tesis se puede desdoblar en dos grandes líneas: primero, el diseño de métodos centrados en la visualización y basados en la inteligencia artificial para el análisis de los datos secuenciales correspondientes a los GPCRs y, segundo, la aplicación de los métodos desarrollados a problemas de farmacoproteómica tales como la subtipificación de GPCRs y el análisis de proteinas no-alineadas

DNA Sequence Classification: It’s Easier Than You Think: An open-source k-mer based machine learning tool for fast and accurate classification of a variety of genomic datasets

Supervised classification of genomic sequences is a challenging, well-studied problem with a variety of important applications. We propose an open-source, supervised, alignment-free, highly general method for sequence classification that operates on k-mer proportions of DNA sequences. This method was implemented in a fully standalone general-purpose software package called Kameris, publicly available under a permissive open-source license. Compared to competing software, ours provides key advantages in terms of data security and privacy, transparency, and reproducibility. We perform a detailed study of its accuracy and performance on a wide variety of classification tasks, including virus subtyping, taxonomic classification, and human haplogroup assignment. We demonstrate the success of our method on whole mitochondrial, nuclear, plastid, plasmid, and viral genomes, as well as randomly sampled eukaryote genomes and transcriptomes. Further, we perform head-to-head evaluations on the tasks of HIV-1 virus subtyping and bacterial taxonomic classification with a number of competing state-of-the-art software solutions, and show that we match or exceed all other tested software in terms of accuracy and speed

Testing robustness of relative complexity measure method constructing robust phylogenetic trees for \u3ci\u3eGalanthus\u3c/i\u3e L. Using the relative complexity measure

Background: Most phylogeny analysis methods based on molecular sequences use multiple alignment where the quality of the alignment, which is dependent on the alignment parameters, determines the accuracy of the resulting trees. Different parameter combinations chosen for the multiple alignment may result in different phylogenies. A new non-alignment based approach, Relative Complexity Measure (RCM), has been introduced to tackle this problem and proven to work in fungi and mitochondrial DNA. Result: In this work, we present an application of the RCM method to reconstruct robust phylogenetic trees using sequence data for genus Galanthus obtained from different regions in Turkey. Phylogenies have been analyzed using nuclear and chloroplast DNA sequences. Results showed that, the tree obtained from nuclear ribosomal RNA gene sequences was more robust, while the tree obtained from the chloroplast DNA showed a higher degree of variation. Conclusions: Phylogenies generated by Relative Complexity Measure were found to be robust and results of RCM were more reliable than the compared techniques. Particularly, to overcome MSA-based problems, RCM seems to be a reasonable way and a good alternative to MSA-based phylogenetic analysis. We believe our method will become a mainstream phylogeny construction method especially for the highly variable sequence families where the accuracy of the MSA heavily depends on the alignment parameters

ProDis-ContSHC: learning protein dissimilarity measures and hierarchical context coherently for protein-protein comparison in protein database retrieval

<p>Abstract</p> <p>Background</p> <p>The need to retrieve or classify protein molecules using structure or sequence-based similarity measures underlies a wide range of biomedical applications. Traditional protein search methods rely on a pairwise dissimilarity/similarity measure for comparing a pair of proteins. This kind of pairwise measures suffer from the limitation of neglecting the distribution of other proteins and thus cannot satisfy the need for high accuracy of the retrieval systems. Recent work in the machine learning community has shown that exploiting the global structure of the database and learning the contextual dissimilarity/similarity measures can improve the retrieval performance significantly. However, most existing contextual dissimilarity/similarity learning algorithms work in an unsupervised manner, which does not utilize the information of the known class labels of proteins in the database.</p> <p>Results</p> <p>In this paper, we propose a novel protein-protein dissimilarity learning algorithm, ProDis-ContSHC. ProDis-ContSHC regularizes an existing dissimilarity measure <it>d_ij</it>by considering the contextual information of the proteins. The context of a protein is defined by its neighboring proteins. The basic idea is, for a pair of proteins (<it>i</it>, <it>j</it>), if their context <inline-formula><m:math xmlns:m="http://www.w3.org/1998/Math/MathML" name="1471-2105-13-S7-S2-i1"><m:mi mathvariant="script">N</m:mi><m:mrow><m:mo class="MathClass-open">(</m:mo><m:mrow><m:mi>i</m:mi></m:mrow><m:mo class="MathClass-close">)</m:mo></m:mrow></m:math></inline-formula> and <inline-formula><m:math xmlns:m="http://www.w3.org/1998/Math/MathML" name="1471-2105-13-S7-S2-i2"><m:mi mathvariant="script">N</m:mi><m:mrow><m:mo class="MathClass-open">(</m:mo><m:mrow><m:mi>j</m:mi></m:mrow><m:mo class="MathClass-close">)</m:mo></m:mrow></m:math></inline-formula> is similar to each other, the two proteins should also have a high similarity. We implement this idea by regularizing <it>d_ij</it>by a factor learned from the context <inline-formula><m:math xmlns:m="http://www.w3.org/1998/Math/MathML" name="1471-2105-13-S7-S2-i3"><m:mi mathvariant="script">N</m:mi><m:mrow><m:mo class="MathClass-open">(</m:mo><m:mrow><m:mi>i</m:mi></m:mrow><m:mo class="MathClass-close">)</m:mo></m:mrow></m:math></inline-formula> and <inline-formula><m:math xmlns:m="http://www.w3.org/1998/Math/MathML" name="1471-2105-13-S7-S2-i4"><m:mi mathvariant="script">N</m:mi><m:mrow><m:mo class="MathClass-open">(</m:mo><m:mrow><m:mi>j</m:mi></m:mrow><m:mo class="MathClass-close">)</m:mo></m:mrow></m:math></inline-formula>.</p> <p>Moreover, we divide the context to hierarchial sub-context and get the contextual dissimilarity vector for each protein pair. Using the class label information of the proteins, we select the relevant (a pair of proteins that has the same class labels) and irrelevant (with different labels) protein pairs, and train an SVM model to distinguish between their contextual dissimilarity vectors. The SVM model is further used to learn a supervised regularizing factor. Finally, with the new <b>S</b>upervised learned <b>Dis</b>similarity measure, we update the <b>Pro</b>tein <b>H</b>ierarchial <b>Cont</b>ext <b>C</b>oherently in an iterative algorithm--<b>ProDis-ContSHC</b>.</p> <p>We test the performance of ProDis-ContSHC on two benchmark sets, i.e., the ASTRAL 1.73 database and the FSSP/DALI database. Experimental results demonstrate that plugging our supervised contextual dissimilarity measures into the retrieval systems significantly outperforms the context-free dissimilarity/similarity measures and other unsupervised contextual dissimilarity measures that do not use the class label information.</p> <p>Conclusions</p> <p>Using the contextual proteins with their class labels in the database, we can improve the accuracy of the pairwise dissimilarity/similarity measures dramatically for the protein retrieval tasks. In this work, for the first time, we propose the idea of supervised contextual dissimilarity learning, resulting in the ProDis-ContSHC algorithm. Among different contextual dissimilarity learning approaches that can be used to compare a pair of proteins, ProDis-ContSHC provides the highest accuracy. Finally, ProDis-ContSHC compares favorably with other methods reported in the recent literature.</p

Contribution à l'analyse des séquences de protéines similarité, clustering et alignement

La prédiction des fonctions biologiques des protéines est primordiale en biologie cellulaire. On peut comprendre facilement tout l'enjeu de pouvoir différencier efficacement les protéines par leurs fonctions, quand on sait que ceci peut rendre possible la réparation des protéines anormales causants des maladies, ou du moins corriger ou améliorer leurs fonctions. Les méthodes expérimentales, basées sur la structure tridimensionnelle des protéines sont les plus fiables pour la prédiction des fonctions biologiques des protéines. Néanmoins, elles sont souvent coûteuses en temps et en ressources, et ne permettent pas de traiter de grands nombres de protéines. Il existe toutefois des algorithmes qui permettent aux biologistes d'arriver à de bons résultats de prédictions en utilisant des moyens beaucoup moins coûteux. Le plus souvent, ces algorithmes sont basés sur la similarité, le clustering, et l'alignement. Cependant, les algorithmes qui sont basés sur la similarité et le clustering utilisent souvent l'alignement des séquences et ne sont donc pas efficaces sur les protéines non alignables. Et lorsqu'ils ne sont pas basés sur l 'alignement, ces algorithmes utilisent souvent des approches qui ne tiennent pas compte de l'aspect biologique des séquences de protéines. D'autre part, l'efficacité des algorithmes d'alignements dépend souvent de la nature structurelle des protéines, ce qui rend difficile le choix de l'algorithme à utiliser quand la structure est inconnue. Par ailleurs, les algorithmes d'alignement ignorent les divergences entre les séquences à aligner, ce qui contraint souvent les biologistes à traiter manuellement les séquences à aligner, une tâche qui n'est pas toujours possible en pratique. Dans cette thèse nous présentons un ensemble de nouveaux algorithmes que nous avons conçus pour l'analyse des séquences de protéines. Dans le premier chapitre, nous présentons CLUSS, le premier algorithme de clustering capable de traiter des séquences de protéines non-alignables. Dans le deuxième chapitre, nous présentons CLUSS2 une version améliorée de CLUSS, capable de traiter de plus grands ensembles de protéines avec plus de de fonctions biologiques. Dans le troisième chapitre, nous présentons SCS, une nouvelle mesure de similarité capable de traiter efficacement non seulement les séquences de protéines mais aussi plusieurs types de séquences catégoriques. Dans le dernier chapitre, nous présentons ALIGNER, un algorithme d'alignement, efficace sur les séquences de protéines indépendamment de leurs types de structures. De plus, ALIGNER est capable de détecter automatiquement, parmi les protéines à aligner, les groupes de protéines dont l'alignement peut révéler d'importantes propriétés biochimiques structurelles et fonctionnelles, et cela sans faire appel à l'utilisateur