10 research outputs found

    Dynamic design: manipulation of millisecond timescale motions on the energy landscape of cyclophilin A

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    Proteins need to interconvert between many conformations in order to function, many of which are formed transiently, and sparsely populated. Particularly when the lifetimes of these states approach the millisecond timescale, identifying the relevant structures and the mechanism by which they interconvert remains a tremendous challenge. Here we introduce a novel combination of accelerated MD (aMD) simulations and Markov state modelling (MSM) to explore these ‘excited’ conformational states. Applying this to the highly dynamic protein CypA, a protein involved in immune response and associated with HIV infection, we identify five principally populated conformational states and the atomistic mechanism by which they interconvert. A rational design strategy predicted that the mutant D66A should stabilise the minor conformations and substantially alter the dynamics, whereas the similar mutant H70A should leave the landscape broadly unchanged. These predictions are confirmed using CPMG and R1ρ solution state NMR measurements. By efficiently exploring functionally relevant, but sparsely populated conformations with millisecond lifetimes in silico, our aMD/MSM method has tremendous promise for the design of dynamic protein free energy landscapes for both protein engineering and drug discovery

    Dynamic Profiling of β-Coronavirus 3CL Mpro Protease Ligand-Binding Sites

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    β-coronavirus (CoVs) alone has been responsible for three major global outbreaks in the 21st century. The current crisis has led to an urgent requirement to develop therapeutics. Even though a number of vaccines are available, alternative strategies targeting essential viral components are required as a backup against the emergence of lethal viral variants. One such target is the main protease (Mpro) that plays an indispensable role in viral replication. The availability of over 270 Mpro X-ray structures in complex with inhibitors provides unique insights into ligand-protein interactions. Herein, we provide a comprehensive comparison of all nonredundant ligand-binding sites available for SARS-CoV2, SARS-CoV, and MERS-CoV Mpro. Extensive adaptive sampling has been used to investigate structural conservation of ligand-binding sites using Markov state models (MSMs) and compare conformational dynamics employing convolutional variational auto-encoder-based deep learning. Our results indicate that not all ligand-binding sites are dynamically conserved despite high sequence and structural conservation across β-CoV homologs. This highlights the complexity in targeting all three Mpro enzymes with a single pan inhibitor

    Machine Learning for Molecular Dynamics on Long Timescales

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    Molecular dynamics (MD) simulation is widely used to analyze the properties of molecules and materials. Most practical applications, such as comparison with experimental measurements, designing drug molecules, or optimizing materials, rely on statistical quantities, which may be prohibitively expensive to compute from direct long-time MD simulations. Classical machine learning (ML) techniques have already had a profound impact on the field, especially for learning low-dimensional models of the long-time dynamics and for devising more efficient sampling schemes for computing long-time statistics. Novel ML methods have the potential to revolutionize long timescale MD and to obtain interpretable models. ML concepts such as statistical estimator theory, end-to-end learning, representation learning, and active learning are highly interesting for the MD researcher and will help to develop new solutions to hard MD problems. With the aim of better connecting the MD and ML research areas and spawning new research on this interface, we define the learning problems in long timescale MD, present successful approaches, and outline some of the unsolved ML problems in this application field
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