The R403Q Myosin Mutation Implicated in Familial Hypertrophic Cardiomyopathy Causes Disorder at the Actomyosin Interface

Mutations in virtually all of the proteins comprising the cardiac muscle sarcomere have been implicated in causing Familial Hypertrophic Cardiomyopathy (FHC). Mutations in the beta-myosin heavy chain (MHC) remain among the most common causes of FHC, with the widely studied R403Q mutation resulting in an especially severe clinical prognosis. In vitro functional studies of cardiac myosin containing the R403Q mutation have revealed significant changes in enzymatic and mechanical properties compared to wild-type myosin. It has been proposed that these molecular changes must trigger events that ultimately lead to the clinical phenotype.Here we examine the structural consequences of the R403Q mutation in a recombinant smooth muscle myosin subfragment (S1), whose kinetic features have much in common with slow beta-MHC. We obtained three-dimensional reconstructions of wild-type and R403Q smooth muscle S1 bound to actin filaments in the presence (ADP) and absence (apo) of nucleotide by electron cryomicroscopy and image analysis. We observed that the mutant S1 was attached to actin at highly variable angles compared to wild-type reconstructions, suggesting a severe disruption of the actin-myosin interaction at the interface.These results provide structural evidence that disarray at the molecular level may be linked to the histopathological myocyte disarray characteristic of the diseased state

Studies of the structure and growth mode of dotriacontane films by synchrotron x-ray scattering and molecular dynamics simulations

doi: 10.1088/0953-8984/16/29/005We report on synchrotron x-ray scattering experiments and molecular dynamics simulations of the structure and growth mode of dotriacontane (n-C32H66 or C32) films adsorbed on Ag(111) and SiO2-coated Si(100) substrates. On the SiO2 surface, the x-ray measurements confirm a structural model of the solid film inferred from high-resolution ellipsometry measurements in which one or two layers of C32 adsorb with the long axis of the molecule oriented parallel to the interface followed by a monolayer in which the molecules have a perpendicular orientation. At higher C32 coverages, preferentially oriented bulk particles nucleate, consistent with a Stranski-Krastanov growth mode. On the Ag(111) surface, we again observe one or two layers of the 'parallel' film but no evidence of the perpendicular monolayer before nucleation of the preferentially oriented bulk particles. We compare the experimentally observed structures with molecular dynamics simulations of a multilayer film of the homologous C24 molecule.This work was support by US National Science Foundation under Grant Nos. DMR-9802476 and DMR-0109057. The Midwest Universities Collaborative Access Team (MUCAT) sector at the Advanced Photon Source (APS) is supported by the US Department of Energy (DOE), Office of Science, Office of Basic Energy Sciences (BES), through Ames Laboratory under ContractNo.W-7405-Eng-82. Use of the APS was supported by the DOE BES under Contract No. W-31-109-ENG-38

The structure of the C-terminal actin-binding domain of talin

Talin is a large dimeric protein that couples integrins to cytoskeletal actin. Here, we report the structure of the C-terminal actin-binding domain of talin, the core of which is a five-helix bundle linked to a C-terminal helix responsible for dimerisation. The NMR structure of the bundle reveals a conserved surface-exposed hydrophobic patch surrounded by positively charged groups. We have mapped the actin-binding site to this surface and shown that helix 1 on the opposite side of the bundle negatively regulates actin binding. The crystal structure of the dimerisation helix reveals an antiparallel coiled-coil with conserved residues clustered on the solvent-exposed face. Mutagenesis shows that dimerisation is essential for filamentous actin (F-actin) binding and indicates that the dimerisation helix itself contributes to binding. We have used these structures together with small angle X-ray scattering to derive a model of the entire domain. Electron microscopy provides direct evidence for binding of the dimer to F-actin and indicates that it binds to three monomers along the long-pitch helix of the actin filament

Nanoscale Observation of Alkane Delayering

Noncontact Atomic Force Microscopy and synchrotron x-ray scattering measurements on dotriacontane (n-C32H66 or C32) films adsorbed on SiO2-coated Si(100) wafers reveal a narrow temperature range near the bulk C32 melting point Tb in which a monolayer phase of C32 molecules oriented perpendicular to surface is stable. This monolayer phase undergoes a delayering transition to a three-dimensional (3D) fluid phase on heating to just above Tb and to a solid 3D phase on cooling below Tb. An equilibrium phase diagram provides a useful framework for interpreting the unusual spreading and receding of the monolayer observed in transitions to and from the respective 3D phases.Comment: 13 pages, 3 figure

A lattice gas model of II-VI(001) semiconductor surfaces

We introduce an anisotropic two-dimensional lattice gas model of metal terminated II-IV(001) seminconductor surfaces. Important properties of this class of materials are represented by effective NN and NNN interactions, which result in the competition of two vacancy structures on the surface. We demonstrate that the experimentally observed c(2x2)-(2x1) transition of the CdTe(001) surface can be understood as a phase transition in thermal equilbrium. The model is studied by means of transfer matrix and Monte Carlo techniques. The analysis shows that the small energy difference of the competing reconstructions determines to a large extent the nature of the different phases. Possible implications for further experimental research are discussed.Comment: 7 pages, 2 figure

The generalized 3-edge-connectivity of lexicographic product graphs

The generalized $k$-edge-connectivity $\lambda_k(G)$ of a graph $G$ is a generalization of the concept of edge-connectivity. The lexicographic product of two graphs $G$ and $H$, denoted by $G\circ H$, is an important graph product. In this paper, we mainly study the generalized 3-edge-connectivity of $G \circ H$, and get upper and lower bounds of $\lambda_3(G \circ H)$. Moreover, all bounds are sharp.Comment: 14 page

A superconducting-nanowire 3-terminal electronic device

In existing superconducting electronic systems, Josephson junctions play a central role in processing and transmitting small-amplitude electrical signals. However, Josephson-junction-based devices have a number of limitations including: (1) sensitivity to magnetic fields, (2) limited gain, (3) inability to drive large impedances, and (4) difficulty in controlling the junction critical current (which depends sensitively on sub-Angstrom-scale thickness variation of the tunneling barrier). Here we present a nanowire-based superconducting electronic device, which we call the nanocryotron (nTron), that does not rely on Josephson junctions and can be patterned from a single thin film of superconducting material with conventional electron-beam lithography. The nTron is a 3-terminal, T-shaped planar device with a gain of ~20 that is capable of driving impedances of more than 100 k{\Omega}, and operates in typical ambient magnetic fields at temperatures of 4.2K. The device uses a localized, Joule-heated hotspot formed in the gate to modulate current flow in a perpendicular superconducting channel. We have characterized the nTron, matched it to a theoretical framework, and applied it both as a digital logic element in a half-adder circuit, and as a digital amplifier for superconducting nanowire single-photon detectors pulses. The nTron has immediate applications in classical and quantum communications, photon sensing and astronomy, and its performance characteristics make it compatible with existing superconducting technologies. Furthermore, because the hotspot effect occurs in all known superconductors, we expect the design to be extensible to other materials, providing a path to digital logic, switching, and amplification in high-temperature superconductors

The “Connection” Between HIV Drug Resistance and RNase H

Currently, nucleoside reverse transcriptase inhibitors (NRTIs) and nonnucleoside reverse transcriptase inhibitors (NNRTIs) are two classes of antiretroviral agents that are approved for treatment of HIV-1 infection. Since both NRTIs and NNRTIs target the polymerase (pol) domain of reverse transcriptase (RT), most genotypic analysis for drug resistance is limited to the first ∼300 amino acids of RT. However, recent studies have demonstrated that mutations in the C-terminal domain of RT, specifically the connection subdomain and RNase H domain, can also increase resistance to both NRTIs and NNRTIs. In this review we will present the potential mechanisms by which mutations in the C-terminal domain of RT influence NRTI and NNRTI susceptibility, summarize the prevalence of the mutations in these regions of RT identified to date, and discuss their importance to clinical drug resistance