Hang Yu

Molecular dynamics simulations of large macromolecular complexes.

Connecting dynamics to structural data from diverse experimental sources, molecular dynamics simulations permit the exploration of biological phenomena in unparalleled detail. Advances in simulations are moving the atomic resolution descriptions of biological systems into the million-to-billion atom regime, in which numerous cell functions reside. In this opinion, we review the progress, driven by large-scale molecular dynamics simulations, in the study of viruses, ribosomes, bioenergetic systems, and other diverse applications. These examples highlight the utility of molecular dynamics simulations in the critical task of relating atomic detail to the function of supramolecular complexes, a task that cannot be achieved by smaller-scale simulations or existing experimental approaches alone.

Membrane Sculpting by F-BAR Domains Studied by Molecular Dynamics Simulations

Interplay between cellular membranes and their peripheral proteins drives many processes in eukaryotic cells. Proteins of the Bin/Amphiphysin/Rvs (BAR) domain family, in particular, play a role in cellular morphogenesis, for example curving planar membranes into tubular membranes. However, it is still unclear how F-BAR domain proteins act on membranes. Electron microscopy revealed that, in vitro, F-BAR proteins form regular lattices on cylindrically deformed membrane surfaces. Using all-atom and coarse-grained (CG) molecular dynamics simulations, we show that such lattices, indeed, induce tubes of observed radii. A 250 ns all-atom simulation reveals that F-BAR domain curves membranes via the so-called scaffolding mechanism. Plasticity of the F-BAR domain permits conformational change in response to membrane interaction, via partial unwinding of the domains 3-helix bundle structure. A CG simulation covering more than 350 µs provides a dynamic picture of membrane tubulation by lattices of F-BAR domains. A series of CG simulations identified the optimal lattice type for membrane sculpting, which matches closely the lattices seen through cryo-electron microscopy.

Molecular orientation studies by pulsed electron-electron double resonance experiments

Pulsed electron-electron double resonance (PELDOR) has proven to be a valuable tool to measure the distribution of long range distances in noncrystalline macromolecules. These experiments commonly use nitroxide spin labels as paramagnetic markers that are covalently attached to the macromolecule at specific positions. Unless these spin labels are flexible in such a manner that they exhibit an almost random orientation, the PELDOR signals will-apart from the interspin distance-also depend on the orientation of the spin labels. This effect needs to be considered in the analysis of PELDOR signals and can, moreover, be used to obtain additional information on the structure of the molecule under investigation. In this work, we demonstrate that the PELDOR signal can be represented as a convolution of a kernel function containing the distance distribution function and an orientation intensity function. The following strategy is proposed to obtain both functions from the experimental data. In a first step, the distance distribution function is estimated by the Tikhonov regularization, using the average over all PELDOR time traces with different frequency offsets and neglecting angular correlations of the spin labels. Second, the convolution relation is employed to determine the orientation intensity function, using again the Tikhonov regularization. Adopting small nitroxide biradical molecules as simple examples, it is shown that the approach works well and is internally consistent. Furthermore, independent molecular dynamics simulations are performed and used to calculate PELDOR signals, distance distributions, and orientational intensity functions. The calculated and experimental results are found to be in excellent overall agreement.

Molecular insights into the membrane-associated phosphatidylinositol 4-kinase IIα.

Phosphatidylinositol 4-kinase IIα (PI4KIIα), a membrane-associated PI kinase, plays a central role in cell signalling and trafficking. Its kinase activity critically depends on palmitoylation of its cysteine-rich motif (-CCPCC-) and is modulated by the membrane environment. Lack of atomic structure impairs our understanding of the mechanism regulating kinase activity. Here we present the crystal structure of human PI4KIIα in ADP-bound form. The structure identifies the nucleotide-binding pocket that differs notably from that found in PI3Ks. Two structural insertions, a palmitoylation insertion and an RK-rich insertion, endow PI4KIIα with the ‘integral’ membrane-binding feature. Molecular dynamics simulations, biochemical and mutagenesis studies reveal that the palmitoylation insertion, containing an amphipathic helix, contributes to the PI-binding pocket and anchors PI4KIIα to the membrane, suggesting that fluctuation of the palmitoylation insertion affects PI4KIIα’s activity. We conclude from our results that PI4KIIα’s activity is regulated indirectly through changes in the membrane environment.

Molecular dynamics simulations demonstrate the regulation of DNA-DNA attraction by H4 histone tail acetylations and mutations.

The positively charged N‐terminal histone tails play a crucial role in chromatin compaction and are important modulators of DNA transcription, recombination, and repair. The detailed mechanism of the interaction of histone tails with DNA remains elusive. To model the unspecific interaction of histone tails with DNA, all‐atom molecular dynamics (MD) simulations were carried out for systems of four DNA 22‐mers in the presence of 20 or 16 short fragments of the H4 histone tail (variations of the 16–23 a. a. KRHRKVLR sequence, as well as the unmodified fragment a. a.13–20, GGAKRHRK). This setup with high DNA concentration, explicit presence of DNA‐DNA contacts, presence of unstructured cationic peptides (histone tails) and K+ mimics the conditions of eukaryotic chromatin. A detailed account of the DNA interactions with the histone tail fragments, K+ and water is presented. Furthermore, DNA structure and dynamics and its interplay with the histone tail fragments binding are analysed. The charged side chains of the lysines and arginines play major roles in the tail‐mediated DNA‐DNA attraction by forming bridges and by coordinating to the phosphate groups and to the electronegative sites in the minor groove. Binding of all species to DNA is dynamic. The structure of the unmodified fully‐charged H4 16–23 a.a. fragment KRHRKVLR is dominated by a stretched conformation. The H4 tail a. a. fragment GGAKRHRK as well as the H4 Lys16 acetylated fragment are highly flexible. The present work allows capturing typical features of the histone tail‐counterion‐DNA structure, interaction and dynamics.

Transient β -hairpin formation in α -synuclein monomer revealed by coarse-grained molecular dynamics simulation

Parkinsons disease, originating from the intrinsically disordered peptide α-synuclein, is a common neurodegenerative disorder that affects more than 5% of the population above age 85. It remains unclear how α-synuclein monomers undergo conformational changes leading to aggregation and formation of fibrils characteristic for the disease. In the present study, we perform molecular dynamics simulations (over 180 μs in aggregated time) using a hybrid-resolution model, Proteins with Atomic details in Coarse-grained Environment (PACE), to characterize in atomic detail structural ensembles of wild type and mutant monomeric α-synuclein in aqueous solution. The simulations reproduce structural properties of α-synuclein characterized in experiments, such as secondary structure content, long-range contacts, chemical shifts, and (3)J(HNHCα )-coupling constants. Most notably, the simulations reveal that a short fragment encompassing region 38-53, adjacent to the non-amyloid-β component region, exhibits a high probability of forming a β-hairpin; this fragment, when isolated from the remainder of α-synuclein, fluctuates frequently into its β-hairpin conformation. Two disease-prone mutations, namely, A30P and A53T, significantly accelerate the formation of a β-hairpin in the stated fragment. We conclude that the formation of a β-hairpin in region 38-53 is a key event during α-synuclein aggregation. We predict further that the G47V mutation impedes the formation of a turn in the β-hairpin and slows down β-hairpin formation, thereby retarding α-synuclein aggregation.

A time course of orchestrated endophilin action in sensing, bending, and stabilizing curved membranes

Endophilin is a founding member of the membrane-bending protein family. It promotes rapid endocytosis on a milliseconds-to-seconds time scale. The short duration of rapid endocytosis requires endophilin to act quickly. High-resolution kinetic measurements show the sequential action that endophilin takes to quickly deform membranes.

Influence of Nitroxide Spin Labels on RNA Structure: A Molecular Dynamics Simulation Study.

Pulsed electron double resonance (PELDOR) experiments on oligonucleotides provide a distance ruler that allows the measurement of nanometer distances accurately. The technique requires attachment of nitroxide spin labels to the nucleotides, which may possibly perturb its conformation. To study to what extent nitroxide spin labels may affect RNA structure, all-atom molecular dynamics simulations in explicit solvent are performed for six double-labeled RNA duplexes. A new parametrization of the force field for the nitroxide spin label is developed, which leads to intramolecular distances that are in good agreement with experimental results. Comparison of the results for spin-labeled and unlabeled RNA reveals that the conformational effect of the spin label depends significantly on whether the spin label is attached to the major or the minor groove of RNA. While major-groove spin labeling may to some extent affect the conformation of nearby base pairs, minor-groove spin labeling has the advantage of mostly preserving the RNA conformation.

How Synaptotagmin I, N-BAR and F-BAR Domains Generate Membrane Curvature

Protein-induced membrane curving governs many cellular processes, including cell division, growth and cell-cell communication. In order to unravel the mechanism for how membrane curvature occurs driven by the proteins, one needs to have detailed molecular pictures on the key membrane-protein interactions and protein-protein interactions. Computer simulation serves as a great tool in providing such molecular pictures and thus in rationalizing how proteins curve membrane. For example, it is known that neurotransmitter release involves a Ca++ ion-regulated fusion process between synaptic vesicles and the presynaptic membrane, but little is known for how such small ions function. With the help of computer simulation, we realized the Ca++-sensor protein, synaptotagmin I, undergoes a conformational transition after binding to both the Ca++ and membrane that differs from the membrane-free crystal/NMR structure. The new synaptotagmin conformation has its C-terminal helix to interact with the presynaptic membrane, and thereby, causes the presynaptic membrane to curve and facilitates membrane-vesicle fusion. Another example in showing the power of computer simulation lies in resolving how N-BAR and F-BAR domains sculpt a flat membrane. As these domains form a lattice in the sculpting process, we identified the optimal lattice type and key protein-protein interactions within the lattice. The whole process of membrane tubulation from a flat membrane was resolved from the simulations, and agreements were found between the lattice structures observed via cryo-electron microscopy and the simulations.