Section: Research Program
Classifying and Mining Protein Structures and Protein Interactions
The scientific discovery process is very often based on cycles of measurement, classification, and generalisation. It is easy to argue that this is especially true in the biological sciences. The proteins that exist today represent the molecular product of some three billion years of evolution. Therefore, comparing protein sequences and structures is important for understanding their functional and evolutionary relationships , . There is now overwhelming evidence that all living organisms and many biological processes share a common ancestry in the tree of life. Historically, much of bioinformatics research has focused on developing mathematical and statistical algorithms to process, analyse, annotate, and compare protein and DNA sequences because such sequences represent the primary form of information in biological systems. However, there is growing evidence that structure-based methods can help to predict networks of protein-protein interactions (PPIs) with greater accuracy than those which do not use structural evidence , . Therefore, developing techniques which can mine knowledge of protein structures and their interactions is an important way to enhance our knowledge of biology .
Quantifying Structural Similarity
Often, proteins may be divided into modular sub-units called domains, which can be associated with specific biological functions. Thus, a protein domain may be considered as the evolutionary unit of biological structure and function . However, while it is well known that the 3D structures of protein domains are often more evolutionarily conserved than their one-dimensional (1D) amino acid sequences, comparing 3D structures is much more difficult than comparing 1D sequences. However, until recently, most evolutionary studies of proteins have compared and clustered 1D amino acid and nucleotide sequences rather than 3D molecular structures.
A pre-requisite for the accurate comparison of protein structures is to have a reliable method for quantifying the structural similarity between pairs of proteins. We recently developed a new protein structure alignment program called Kpax which combines an efficient dynamic programming based scoring function with a simple but novel Gaussian representation of protein backbone shape . This means that we can now quantitatively compare 3D protein domains at a similar rate to throughput to conventional protein sequence comparison algorithms. We recently compared Kpax with a large number of other structure alignment programs, and we found Kpax to be the fastest and amongst the most accurate, in a CATH family recognition test . The latest version of Kpax  can calculate multiple flexible alignments, and thus promises to avoid such issues when comparing more distantly related protein folds and fold families.
Formalising and Exploiting Domain Knowledge
Concerning protein structure classification, we aim to explore novel classification paradigms to circumvent the problems encountered with existing hierarchical classifications of protein folds and domains. In particular it will be interesting to set up fuzzy clustering methods taking advantage of our previous work on gene functional classification , but instead using Kpax domain-domain similarity matrices. A non-trivial issue with fuzzy clustering is how to handle similarity rather than mathematical distance matrices, and how to find the optimal number of clusters, especially when using a non-Euclidean similarity measure. We will adapt the algorithms and the calculation of quality indices to the Kpax similarity measure. More fundamentally, it will be necessary to integrate this classification step in the more general process leading from data to knowledge called Knowledge Discovery in Databases (KDD) .
Another example where domain knowledge can be useful is during result interpretation: several sources of knowledge have to be used to explicitly characterise each cluster and to help decide its validity. Thus, it will be useful to be able to express data models, patterns, and rules in a common formalism using a defined vocabulary for concepts and relationships. Existing approaches such as the Molecular Interaction (MI) format  developed by the Human Genome Organization (HUGO) mostly address the experimental wet lab aspects leading to data production and curation . A different point of view is represented in the Interaction Network Ontology (INO), a community-driven ontology that aims to standardise and integrate data on interaction networks and to support computer-assisted reasoning . However, this ontology does not integrate basic 3D concepts and structural relationships. Therefore, extending such formalisms and symbolic relationships will be beneficial, if not essential, when classifying the 3D shapes of proteins at the domain family level.
3D Protein Domain Annotation and Shape Mining
A widely used collection of protein domain families is “Pfam” , constructed from multiple alignments of protein sequences. Integrating domain-domain similarity measures with knowledge about domain binding sites, as introduced by us in our KBDOCK approach , , can help in selecting interesting subsets of domain pairs before clustering. Thanks to our KBDOCK and Kpax projects, we already have a rich set of tools with which we can start to process and compare all known protein structures and PPIs according to their component Pfam domains. Linking this new classification to the latest “SIFTS” (Structure Integration with Function, Taxonomy and Sequence)  functional annotations between standard UniProt (http://www.uniprot.org/ sequence identifiers and protein structures from the Protein Data Bank (PDB)  could then provide a useful way to discover new structural and functional relationships which are difficult to detect in existing classification schemes such as CATH or SCOP. As part of the thesis project of Seyed Alborzi, we developed a recommender-based data mining technique to associate enzyme classification code numbers with Pfam domains using our recently developed EC-DomainMiner program . We subsequently generalised this approach as a tripartite graph mining method for inferring associations between different protein annotation sources, which we call “CODAC” (for COmputational Discovery of Direct Associations using Common Neighbours). A first paper on CODAC was presented at IWBBIO-2017 , and a full paper has recently been accepted by BMC Bioinformatics .
Protein Function Annotation
Knowledge of the functional properties of proteins can shed considerable light on how they might interact. However, huge numbers of protein sequences in public databases lack any functional annotation, and the annotation of sequences in such databases is a highly challenging problem. We are developing graph-based and machine learning techniques to annotate automatically the available unannotated sequences in such databases with functional properties such as EC numbers and Gene Ontology (GO) terms. Even if the 3D structures of proteins are unknown, it is natural to suppose that their sequences may be related to each other by the domains, domain families, and super-families that they share. In the frame of the PhD project of Bishnu Sarker, we recently developed a novel graph-based approach called GrAPFI for the automatic functional annotation of protein sequences based on these principles in order to transfer annotations from expert-reviewed sequences to unreviewed sequences in the UniProtKB databases , .