Combining Coarse-Grained Protein Models with Replica-Exchange All-Atom Molecular Dynamics

We describe a combination of all-atom simulations with CABS, a well-established coarse-grained protein modeling tool, into a single multiscale protocol. The simulation method has been tested on the C-terminal beta hairpin of protein G, a model system of protein folding. After reconstructing atomistic details, conformations derived from the CABS simulation were subjected to replica-exchange molecular dynamics simulations with OPLS-AA and AMBER99sb force fields in explicit solvent. Such a combination accelerates system convergence several times in comparison with all-atom simulations starting from the extended chain conformation, demonstrated by the analysis of melting curves, the number of native-like conformations as a function of time and secondary structure propagation. The results strongly suggest that the proposed multiscale method could be an efficient and accurate tool for high-resolution studies of protein folding dynamics in larger systems.Comment: 12 pages, 4 figure

Towards the high-resolution protein structure prediction. Fast refinement of reduced models with all-atom force field

<p>Abstract</p> <p>Background</p> <p>Although experimental methods for determining protein structure are providing high resolution structures, they cannot keep the pace at which amino acid sequences are resolved on the scale of entire genomes. For a considerable fraction of proteins whose structures will not be determined experimentally, computational methods can provide valuable information. The value of structural models in biological research depends critically on their quality. Development of high-accuracy computational methods that reliably generate near-experimental quality structural models is an important, unsolved problem in the protein structure modeling.</p> <p>Results</p> <p>Large sets of structural decoys have been generated using reduced conformational space protein modeling tool CABS. Subsequently, the reduced models were subject to all-atom reconstruction. Then, the resulting detailed models were energy-minimized using state-of-the-art all-atom force field, assuming fixed positions of the alpha carbons. It has been shown that a very short minimization leads to the proper ranking of the quality of the models (distance from the native structure), when the all-atom energy is used as the ranking criterion. Additionally, we performed test on medium and low accuracy decoys built via classical methods of comparative modeling. The test placed our model evaluation procedure among the state-of-the-art protein model assessment methods.</p> <p>Conclusion</p> <p>These test computations show that a large scale high resolution protein structure prediction is possible, not only for small but also for large protein domains, and that it should be based on a hierarchical approach to the modeling protocol. We employed Molecular Mechanics with fixed alpha carbons to rank-order the all-atom models built on the scaffolds of the reduced models. Our tests show that a physic-based approach, usually considered computationally too demanding for large-scale applications, can be effectively used in such studies.</p

Adaptation patterns of winter wheat cultivars in agro-ecological regions

Multi-environment trials are commonly used to assess cultivar adaptation patterns under different environmental conditions and to help make effective cultivar recommendations for growers. An example of a multi-environment trial system used for cultivar recommendations is the Polish Post-registration Variety Testing System. A common approach in cultivar recommendations is to evaluate the adaptability of cultivars across, or for, specific trial locations. However, the locations of the trials and the fields where a farmer will grow a crop are hardly ever in the same place. Therefore, it would be better to group the trial locations into regions and give recommendations for the whole region. The aim of this study is to evaluate the grain yield adaptation patterns of 62 modern winter wheat cultivars in six agro-ecological regions of Poland for two crop management intensities over five growing seasons. The analysis of the grain yield data was performed separately for each intensity using single-stage approaches in linear mixed models. We ascertained that winter wheat yield variability was in the main determined by agro-ecological region and their interactions, and to a small extent by the cultivar effect. Cultivars Sailor and Linus were widely adapted to all agro-ecological regions studied for both crop management intensities. It is highly probable that these two cultivars will obtain high yield in all agro-ecological regions as well as with both crop management intensities studied. We observed high compatibility rankings between locations for both crop management intensities. High compatibility of the cultivar rankings in the trial locations also provides high precision when determining regions

Four-way data analysis within the linear mixed modelling framework

Cultivars have to be evaluated under different crop management systems across agro-ecosystems and years using multi-environment trials (MET) before releasing them to the market. Frequently, data collected in METs are arranged according to cultivar (G), management (M), location, (L) and year (Y) combinations in a four-way G x M x L x Y data table that is highly unbalanced for cultivars across locations and time. Therefore, we present the restricted maximum likelihood method (REML) for linear mixed models (LMM) with a factor analytic variance-covariance matrix for assessing cultivar adaptation to crop management systems and environments based on unbalanced datasets. Such a multi-environmental trial system has been in operation in Poland for winter wheat (Triticum aestivum L.) in the form of the Post-registration Variety Testing System (PVTS). This study aimed to illustrate the use of LMM in the analysis of unbalanced four-way G x M x L x Y data. LMM analysis provided adjusted means of grain yield for 51 winter wheat cultivars bred in different regions in Europe, tested across 18 trial locations and seven consecutive cropping seasons in two crop management intensities. The application of the four-way LMM with a factor analytic variance-covariance matrix is a complementary and effective tool for evaluating the unbalanced G x M x L x Y table. Cultivars tested had different adaptive responses to the Polish agro-ecosystems separately for each of the crop management intensities. Wide adaptation in both crop management systems was exhibited by cultivars Mulan and Jenga bred in Germany

Ranking models of transmembrane β-barrel proteins using Z-coordinate predictions

Motivation: Transmembrane β-barrels exist in the outer membrane of gram-negative bacteria as well as in chloroplast and mitochondria. They are often involved in transport processes and are promising antimicrobial drug targets. Structures of only a few β-barrel protein families are known. Therefore, a method that could automatically generate such models would be valuable. The symmetrical arrangement of the barrels suggests that an approach based on idealized geometries may be successful

Dodging the crisis of folding proteins with knots

Proteins with nontrivial topology, containing knots and slipknots, have the ability to fold to their native states without any additional external forces invoked. A mechanism is suggested for folding of these proteins, such as YibK and YbeA, which involves an intermediate configuration with a slipknot. It elucidates the role of topological barriers and backtracking during the folding event. It also illustrates that native contacts are sufficient to guarantee folding in around 1-2% of the simulations, and how slipknot intermediates are needed to reduce the topological bottlenecks. As expected, simulations of proteins with similar structure but with knot removed fold much more efficiently, clearly demonstrating the origin of these topological barriers. Although these studies are based on a simple coarse-grained model, they are already able to extract some of the underlying principles governing folding in such complex topologies.Comment: 29 pages, 11 figures, 1 tabl

Type II restriction endonuclease R.Eco29kI is a member of the GIY-YIG nuclease superfamily

<p>Abstract</p> <p>Background</p> <p>The majority of experimentally determined crystal structures of Type II restriction endonucleases (REases) exhibit a common PD-(D/E)XK fold. Crystal structures have been also determined for single representatives of two other folds: PLD (R.BfiI) and half-pipe (R.PabI), and bioinformatics analyses supported by mutagenesis suggested that some REases belong to the HNH fold. Our previous bioinformatic analysis suggested that REase R.Eco29kI shares sequence similarities with one more unrelated nuclease superfamily, GIY-YIG, however so far no experimental data were available to support this prediction. The determination of a crystal structure of the GIY-YIG domain of homing endonuclease I-TevI provided a template for modeling of R.Eco29kI and prompted us to validate the model experimentally.</p> <p>Results</p> <p>Using protein fold-recognition methods we generated a new alignment between R.Eco29kI and I-TevI, which suggested a reassignment of one of the putative catalytic residues. A theoretical model of R.Eco29kI was constructed to illustrate its predicted three-dimensional fold and organization of the active site, comprising amino acid residues Y49, Y76, R104, H108, E142, and N154. A series of mutants was constructed to generate amino acid substitutions of selected residues (Y49A, R104A, H108F, E142A and N154L) and the mutant proteins were examined for their ability to bind the DNA containing the Eco29kI site 5'-CCGCGG-3' and to catalyze the cleavage reaction. Experimental data reveal that residues Y49, R104, E142, H108, and N154 are important for the nuclease activity of R.Eco29kI, while H108 and N154 are also important for specific DNA binding by this enzyme.</p> <p>Conclusion</p> <p>Substitutions of residues Y49, R104, H108, E142 and N154 predicted by the model to be a part of the active site lead to mutant proteins with strong defects in the REase activity. These results are in very good agreement with the structural model presented in this work and with our prediction that R.Eco29kI belongs to the GIY-YIG superfamily of nucleases. Our study provides the first experimental evidence for a Type IIP REase that does not belong to the PD-(D/E)XK or HNH superfamilies of nucleases, and is instead a member of the unrelated GIY-YIG superfamily.</p

Optimized Folding Simulations of Protein A

We describe optimized parallel tempering simulations of the 46-residue B-fragment of protein A. Native-like configurations with a root-mean-square deviation of approximately 3A to the experimentally determined structure (Protein Data Bank identifier 1BDD) are found. However, at biologically relevant temperatures such conformations appear with only about 10% frequency in our simulations. Possible short comings in our energy function are discussed.Comment: 6 pages, 8 figure