LOMETS2: improved meta-threading server for fold-recognition and structure-based function annotation for distant-homology proteins.
ABSTRACT: The LOMETS2 server (https://zhanglab.ccmb.med.umich.edu/LOMETS/) is an online meta-threading server system for template-based protein structure prediction. Although the server has been widely used by the community over the last decade, the previous LOMETS server no longer represents the state-of-the-art due to aging of the algorithms and unsatisfactory performance on distant-homology template identification. An extension of the server built on cutting-edge methods, especially techniques developed since the recent CASP experiments, is urgently needed. In this work, we report the recent advancements of the LOMETS2 server, which comprise a number of major new developments, including (i) new state-of-the-art threading programs, including contact-map-based threading approaches, (ii) deep sequence search-based sequence profile construction and (iii) a new web interface design that incorporates structure-based function annotations. Large-scale benchmark tests demonstrated that the integration of the deep profiles and new threading approaches into LOMETS2 significantly improve its structure modeling quality and template detection, where LOMETS2 detected 176% more templates with TM-scores >0.5 than the previous LOMETS server for Hard targets that lacked homologous templates. Meanwhile, the newly incorporated structure-based function prediction helps extend the usefulness of the online server to the broader biological community.
Project description:The COFACTOR web server is a unified platform for structure-based multiple-level protein function predictions. By structurally threading low-resolution structural models through the BioLiP library, the COFACTOR server infers three categories of protein functions including gene ontology, enzyme commission and ligand-binding sites from various analogous and homologous function templates. Here, we report recent improvements of the COFACTOR server in the development of new pipelines to infer functional insights from sequence profile alignments and protein-protein interaction networks. Large-scale benchmark tests show that the new hybrid COFACTOR approach significantly improves the function annotation accuracy of the former structure-based pipeline and other state-of-the-art functional annotation methods, particularly for targets that have no close homology templates. The updated COFACTOR server and the template libraries are available at http://zhanglab.ccmb.med.umich.edu/COFACTOR/.
Project description:Pair-wise residue-residue contacts in proteins can be predicted from both threading templates and sequence-based machine learning. However, most structure modeling approaches only use the template-based contact predictions in guiding the simulations; this is partly because the sequence-based contact predictions are usually considered to be less accurate than that by threading. With the rapid progress in sequence databases and machine-learning techniques, it is necessary to have a detailed and comprehensive assessment of the contact-prediction methods in different template conditions.We develop two methods for protein-contact predictions: SVM-SEQ is a sequence-based machine learning approach which trains a variety of sequence-derived features on contact maps; SVM-LOMETS collects consensus contact predictions from multiple threading templates. We test both methods on the same set of 554 proteins which are categorized into 'Easy', 'Medium', 'Hard' and 'Very Hard' targets based on the evolutionary and structural distance between templates and targets. For the Easy and Medium targets, SVM-LOMETS obviously outperforms SVM-SEQ; but for the Hard and Very Hard targets, the accuracy of the SVM-SEQ predictions is higher than that of SVM-LOMETS by 12-25%. If we combine the SVM-SEQ and SVM-LOMETS predictions together, the total number of correctly predicted contacts in the Hard proteins will increase by more than 60% (or 70% for the long-range contact with a sequence separation > or =24), compared with SVM-LOMETS alone. The advantage of SVM-SEQ is also shown in the CASP7 free modeling targets where the SVM-SEQ is around four times more accurate than SVM-LOMETS in the long-range contact prediction. These data demonstrate that the state-of-the-art sequence-based contact prediction has reached a level which may be helpful in assisting tertiary structure modeling for the targets which do not have close structure templates. The maximum yield should be obtained by the combination of both sequence- and template-based predictions.
Project description:The I-TASSER server (http://zhanglab.ccmb.med.umich.edu/I-TASSER) is an online resource for automated protein structure prediction and structure-based function annotation. In I-TASSER, structural templates are first recognized from the PDB using multiple threading alignment approaches. Full-length structure models are then constructed by iterative fragment assembly simulations. The functional insights are finally derived by matching the predicted structure models with known proteins in the function databases. Although the server has been widely used for various biological and biomedical investigations, numerous comments and suggestions have been reported from the user community. In this article, we summarize recent developments on the I-TASSER server, which were designed to address the requirements from the user community and to increase the accuracy of modeling predictions. Focuses have been made on the introduction of new methods for atomic-level structure refinement, local structure quality estimation and biological function annotations. We expect that these new developments will improve the quality of the I-TASSER server and further facilitate its use by the community for high-resolution structure and function prediction.
Project description:We develop and test a new pipeline in CASP10 to predict protein structures based on an interplay of I-TASSER and QUARK for both free-modeling (FM) and template-based modeling (TBM) targets. The most noteworthy observation is that sorting through the threading template pool using the QUARK-based ab initio models as probes allows the detection of distant-homology templates which might be ignored by the traditional sequence profile-based threading alignment algorithms. Further template assembly refinement by I-TASSER resulted in successful folding of two medium-sized FM targets with >150 residues. For TBM, the multiple threading alignments from LOMETS are, for the first time, incorporated into the ab initio QUARK simulations, which were further refined by I-TASSER assembly refinement. Compared with the traditional threading assembly refinement procedures, the inclusion of the threading-constrained ab initio folding models can consistently improve the quality of the full-length models as assessed by the GDT-HA and hydrogen-bonding scores. Despite the success, significant challenges still exist in domain boundary prediction and consistent folding of medium-size proteins (especially beta-proteins) for nonhomologous targets. Further developments of sensitive fold-recognition and ab initio folding methods are critical for solving these problems.
Project description:We have developed a new COFACTOR webserver for automated structure-based protein function annotation. Starting from a structural model, given by either experimental determination or computational modeling, COFACTOR first identifies template proteins of similar folds and functional sites by threading the target structure through three representative template libraries that have known protein-ligand binding interactions, Enzyme Commission number or Gene Ontology terms. The biological function insights in these three aspects are then deduced from the functional templates, the confidence of which is evaluated by a scoring function that combines both global and local structural similarities. The algorithm has been extensively benchmarked by large-scale benchmarking tests and demonstrated significant advantages compared to traditional sequence-based methods. In the recent community-wide CASP9 experiment, COFACTOR was ranked as the best method for protein-ligand binding site predictions. The COFACTOR sever and the template libraries are freely available at http://zhanglab.ccmb.med.umich.edu/COFACTOR.
Project description:We develop a hierarchical pipeline, ThreaDomEx, for both continuous domain (CD) and discontinuous domain (DCD) structure predictions. Starting from a query sequence, ThreaDomEx first threads it through the PDB to identify multiple structure templates, where a profile of domain conservation score (DC-score) is derived for domain-segment assignment. To further detect DCDs that consist of separated segments along the sequence, a boundary-clustering algorithm is used to refine the DCD-linker locations. In case that the templates do not contain DCDs, a domain-segment assembly process, guided by symmetry comparison, is applied for further DCD detections. ThreaDomEx was tested a set of 1111 proteins and achieved a normalized domain overlap score of 89.3% compared to experimental data, which is significantly higher than other state-of-the-art methods. It also recalls 26.7% of DCDs with 72.7% precision on the proteins for which threading failed to detect any DCDs. The server provides facilities for users to interactively refine the domain models by adjusting DC-score threshold, deleting and adding domain linkers, and assembling domain segments, which are particularly helpful for the hard targets for which current methods have a low accuracy while human-expert knowledge and experimental insights can be used for refining models. ThreaDomEX server is available at http://zhanglab.ccmb.med.umich.edu/ThreaDomEx.
Project description:Most threading methods predict the structure of a protein using only a single template. Due to the increasing number of solved structures, a protein without solved structure is very likely to have more than one similar template structures. Therefore, a natural question to ask is if we can improve modeling accuracy using multiple templates. This article describes a new multiple-template threading method to answer this question. At the heart of this multiple-template threading method is a novel probabilistic-consistency algorithm that can accurately align a single protein sequence simultaneously to multiple templates. Experimental results indicate that our multiple-template method can improve pairwise sequence-template alignment accuracy and generate models with better quality than single-template models even if they are built from the best single templates (P-value <10(-6)) while many popular multiple sequence/structure alignment tools fail to do so. The underlying reason is that our probabilistic-consistency algorithm can generate accurate multiple sequence/template alignments. In another word, without an accurate multiple sequence/template alignment, the modeling accuracy cannot be improved by simply using multiple templates to increase alignment coverage. Blindly tested on the CASP9 targets with more than one good template structures, our method outperforms all other CASP9 servers except two (Zhang-Server and QUARK of the same group). Our probabilistic-consistency algorithm can possibly be extended to align multiple protein/RNA sequences and structures.
Project description:BACKGROUND: Protein inter-residue contacts play a crucial role in the determination and prediction of protein structures. Previous studies on contact prediction indicate that although template-based consensus methods outperform sequence-based methods on targets with typical templates, such consensus methods perform poorly on new fold targets. However, we find out that even for new fold targets, the models generated by threading programs can contain many true contacts. The challenge is how to identify them. RESULTS: In this paper, we develop an integer linear programming model for consensus contact prediction. In contrast to the simple majority voting method assuming that all the individual servers are equally important and independent, the newly developed method evaluates their correlation by using maximum likelihood estimation and extracts independent latent servers from them by using principal component analysis. An integer linear programming method is then applied to assign a weight to each latent server to maximize the difference between true contacts and false ones. The proposed method is tested on the CASP7 data set. If the top L/5 predicted contacts are evaluated where L is the protein size, the average accuracy is 73%, which is much higher than that of any previously reported study. Moreover, if only the 15 new fold CASP7 targets are considered, our method achieves an average accuracy of 37%, which is much better than that of the majority voting method, SVM-LOMETS, SVM-SEQ, and SAM-T06. These methods demonstrate an average accuracy of 13.0%, 10.8%, 25.8% and 21.2%, respectively. CONCLUSION: Reducing server correlation and optimally combining independent latent servers show a significant improvement over the traditional consensus methods. This approach can hopefully provide a powerful tool for protein structure refinement and prediction use.
Project description:Molecular replacement (MR) is one of the most common techniques used for solving the phase problem in X-ray crystal diffraction. The success rate of MR however drops quickly when the sequence identity between query and templates is reduced, while the I-TASSER-MR server is designed to solve the phase problem for proteins that lack close homologous templates. Starting from a sequence, it first generates full-length models using I-TASSER by iterative structural fragment reassembly. A progressive sequence truncation procedure is then used for editing the models based on local variations of the structural assembly simulations. Next, the edited models are submitted to MR-REX to search for optimal placements in the crystal unit-cells through replica-exchange Monte Carlo simulations, with the phasing results used by CNS for final atomic model refinement and selection. The I-TASSER-MR algorithm was tested in large-scale benchmark datasets and solved 36% more targets compared to using the best threading templates. The server takes primary sequence and raw crystal diffraction data as input, with output containing annotated phase information and refined structure models. It also allows users to choose between different methods for setting B-factors and the number of models used for phasing. The online server is freely available at http://zhanglab.ccmb.med.umich.edu/I-TASSER-MR.
Project description:Accurate prediction of atomic-level protein structure is important for annotating the biological functions of protein molecules and for designing new compounds to regulate the functions. Template-based modeling (TBM), which aims to construct structural models by copying and refining the structural frameworks of other known proteins, remains the most accurate method for protein structure prediction. Due to the difficulty in recognizing distant-homology templates, however, the accuracy of TBM decreases rapidly when the evolutionary relationship between the query and template vanishes. In this study, we propose a new method, CEthreader, which first predicts residue-residue contacts by coupling evolutionary precision matrices with deep residual convolutional neural-networks. The predicted contact maps are then integrated with sequence profile alignments to recognize structural templates from the PDB. The method was tested on two independent benchmark sets consisting collectively of 1,153 non-homologous protein targets, where CEthreader detected 176% or 36% more correct templates with a TM-score >0.5 than the best state-of-the-art profile- or contact-based threading methods, respectively, for the Hard targets that lacked homologous templates. Moreover, CEthreader was able to identify 114% or 20% more correct templates with the same Fold as the query, after excluding structures from the same SCOPe Superfamily, than the best profile- or contact-based threading methods. Detailed analyses show that the major advantage of CEthreader lies in the efficient coupling of contact maps with profile alignments, which helps recognize global fold of protein structures when the homologous relationship between the query and template is weak. These results demonstrate an efficient new strategy to combine ab initio contact map prediction with profile alignments to significantly improve the accuracy of template-based structure prediction, especially for distant-homology proteins.