Allosteric regulation of 3CL protease of SARS-CoV-2 and SARS-CoV observed in the crystal structure ensemble

The 3C-like protease (3CLpro) of SARS-CoV-2 is a potential therapeutic target for COVID-19. Importantly, it has an abundance of structural information solved as a complex with various drug candidate compounds. Collecting these crystal structures (83 Protein Data Bank (PDB) entries) together with those of the highly homologous 3CLpro of SARS-CoV (101 PDB entries), we constructed the crystal structure ensemble of 3CLpro to analyze the dynamic regulation of its catalytic function. The structural dynamics of the 3CLpro dimer observed in the ensemble were characterized by the motions of four separate loops (the C-loop, E-loop, H-loop, and Linker) and the C-terminal domain III on the rigid core of the chymotrypsin fold. Among the four moving loops, the C-loop (also known as the oxyanion binding loop) causes the order (active)-disorder (collapsed) transition, which is regulated cooperatively by five hydrogen bonds made with the surrounding residues. The C-loop, E-loop, and Linker constitute the major ligand binding sites, which consist of a limited variety of binding residues including the substrate binding subsites. Ligand binding causes a ligand size dependent conformational change to the E-loop and Linker, which further stabilize the C-loop via the hydrogen bond between the C-loop and E-loop. The T285A mutation from SARS-CoV 3CLpro to SARS-CoV-2 3CLpro significantly closes the interface of the domain III dimer and allosterically stabilizes the active conformation of the C-loop via hydrogen bonds with Ser1 and Gly2; thus, SARS-CoV-2 3CLpro seems to have increased activity relative to that of SARS-CoV 3CLpro.

A method for validating the accuracy of NMR protein structures

We present a method that measures the accuracy of NMR protein structures. It compares random coil index [RCI] against local rigidity predicted by mathematical rigidity theory, calculated from NMR structures [FIRST], using a correlation score (which assesses secondary structure), and an RMSD score (which measures overall rigidity). We test its performance using: structures refined in explicit solvent, which are much better than unrefined structures; decoy structures generated for 89 NMR structures; and conventional predictors of accuracy such as number of restraints per residue, restraint violations, energy of structure, ensemble RMSD, Ramachandran distribution, and clashscore. Restraint violations and RMSD are poor measures of accuracy. Comparisons of NMR to crystal structures show that secondary structure is equally accurate, but crystal structures are typically too rigid in loops, whereas NMR structures are typically too floppy overall. We show that the method is a useful addition to existing measures of accuracy.

PEnBayes: A Multi-Layered Ensemble Approach for Learning Bayesian Network Structure from Big Data

Discovering the Bayesian network (BN) structure from big datasets containing rich causal relationships is becoming increasingly valuable for modeling and reasoning under uncertainties in many areas with big data gathered from sensors due to high volume and fast veracity. Most of the current BN structure learning algorithms have shortcomings facing big data. First, learning a BN structure from the entire big dataset is an expensive task which often ends in failure due to memory constraints. Second, it is quite difficult to select a learner from numerous BN structure learning algorithms to consistently achieve good learning accuracy. Lastly, there is a lack of an intelligent method that merges separately learned BN structures into a well structured BN network. To address these shortcomings, we introduce a novel parallel learning approach called PEnBayes (Parallel Ensemble-based Bayesian network learning). PEnBayes starts with an adaptive data preprocessing phase that calculates the Appropriate Learning Size and intelligently divides a big dataset for fast distributed local structure learning. Then, PEnBayes learns a collection of local BN Structures in parallel using a two-layered weighted adjacent matrix-based structure ensemble method. Lastly, PEnBayes merges the local BN Structures into a global network structure using the structure ensemble method at the global layer. For the experiment, we generate big data sets by simulating sensor data from patient monitoring, transportation, and disease diagnosis domains. The Experimental results show that PEnBayes achieves a significantly improved execution performance with more consistent and stable results compared with three baseline learning algorithms.