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https://www.readbyqxmd.com/read/29779042/combining-theoretical-and-experimental-data-to-decipher-cftr-3d-structures-and-functions
#1
Brice Hoffmann, Ahmad Elbahnsi, Pierre Lehn, Jean-Luc Décout, Fabio Pietrucci, Jean-Paul Mornon, Isabelle Callebaut
Cryo-electron microscopy (cryo-EM) has recently provided invaluable experimental data about the full-length cystic fibrosis transmembrane conductance regulator (CFTR) 3D structure. However, this experimental information deals with inactive states of the channel, either in an apo, quiescent conformation, in which nucleotide-binding domains (NBDs) are widely separated or in an ATP-bound, yet closed conformation. Here, we show that 3D structure models of the open and closed forms of the channel, now further supported by metadynamics simulations and by comparison with the cryo-EM data, could be used to gain some insights into critical features of the conformational transition toward active CFTR forms...
May 19, 2018: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/29775653/atomic-structure-of-a-rationally-engineered-gene-delivery-vector-aav2-5
#2
Matthew Burg, Claire Rosebrough, Lauren M Drouin, Antonette Bennett, Mario Mietzsch, Paul Chipman, Robert McKenna, Duncan Sousa, Mark Potter, Barry Byrne, R Jude Samulski, Mavis Agbandje-McKenna
AAV2.5 represents the first structure-guided in-silico designed Adeno-associated virus (AAV) gene delivery vector. This engineered vector combined the receptor attachment properties of AAV serotype 2 (AAV2) with the muscle tropic properties of AAV1, and exhibited an antibody escape phenotype because of a modified antigenic epitope. To confirm the design, the structure of the vector was determined to a resolution of 2.78 Å using cryo-electron microscopy and image reconstruction. The structure of the major viral protein (VP), VP3, was ordered from residue 219 to 736, as reported for other AAV structures, and the five AAV2...
May 15, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29771342/recent-progress-in-structural-biology-lessons-from-our-research-history
#3
Ryo Nitta, Tsuyoshi Imasaki, Eriko Nitta
The recent 'resolution revolution' in structural analyses of cryo-electron microscopy (cryo-EM) has drastically changed the research strategy for structural biology. In addition to X-ray crystallography and nuclear magnetic resonance spectroscopy, cryo-EM has achieved the structural analysis of biological molecules at near-atomic resolution, resulting in the Nobel Prize in Chemistry 2017. The effect of this revolution has spread within the biology and medical science fields affecting everything from basic research to pharmaceutical development by visualizing atomic structure...
May 16, 2018: Microscopy
https://www.readbyqxmd.com/read/29770216/assessment-of-a-novel-cryoablation-device-for-the-endovascular-treatment-of-cardiac-tachyarrhythmias
#4
John M Baust, Anthony Robilotto, Peter Guerra, Kristi K Snyder, Robert G Van Buskirk, Marc Dubuc, John G Baust
Objectives: Cryoablation is an effective alternative treatment for cardiac arrhythmias offering shortened recovery and reduced side effects. As the use of cryoablation increases, the need for new devices and procedures has emerged. This has been driven by technological limitations including lengthy periods to generate a single lesion (3-5 min), uncertain transmurality, and differential efficacy. Furthermore, due to limited ablation capacity under high heat loads, cryo has had limited success in the treatment of ventricular arrhythmias...
2018: SAGE Open Medicine
https://www.readbyqxmd.com/read/29769723/structure-of-a-volume-regulated-anion-channel-of-the-lrrc8-family
#5
Dawid Deneka, Marta Sawicka, Andy K M Lam, Cristina Paulino, Raimund Dutzler
Volume-regulated anion channels are activated in response to hypotonic stress. These channels are composed of closely related paralogues of the leucine-rich repeat-containing protein 8 (LRRC8) family that co-assemble to form hexameric complexes. Here, using cryo-electron microscopy and X-ray crystallography, we determine the structure of a homomeric channel of the obligatory subunit LRRC8A. This protein conducts ions and has properties in common with endogenous heteromeric channels. Its modular structure consists of a transmembrane pore domain followed by a cytoplasmic leucine-rich repeat domain...
May 16, 2018: Nature
https://www.readbyqxmd.com/read/29769334/reovirus-nonstructural-protein-%C3%AF-ns-acts-as-an-rna-stability-factor-promoting-viral-genome-replication
#6
Paula F Zamora, Liya Hu, Jonathan J Knowlton, Roni M Lahr, Rodolfo A Moreno, Andrea J Berman, B V Venkataram Prasad, Terence S Dermody
Viral nonstructural proteins, which are not packaged into virions, are essential for replication of most viruses. Reovirus, a nonenveloped, double-stranded RNA (dsRNA) virus, encodes three nonstructural proteins that are required for viral replication and dissemination in the host. Reovirus nonstructural protein σNS is a single-stranded RNA (ssRNA)-binding protein that must be expressed in infected cells for production of viral progeny. However, activities of σNS during individual steps of the reovirus replication cycle are poorly understood...
May 16, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29767998/rapid-on-chip-assembly-of-niosomes-batch-versus-continuous-flow-reactors
#7
Sara Garcia-Salinas, Erico Himawan, Gracia Mendoza, Manuel Arruebo, Victor Sebastian Cabeza
The large-scale continuous production of niosomes remains challenging. The inherent drawbacks of batch processes such as large particle polydispersity and reduced batch-to-batch reproducibility are here overcome by using commercially available microfluidic reactors. Compared to the traditional batch-based film hydration method, herein, we demonstrate that it is possible to carry out the homogeneous, large-scale (up to 120mg/min) production of niosomes using two different synthesis techniques (the thin film hydration method and the emulsification technique)...
May 16, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29765610/x-ray-and-cryo-em-structures-of-inhibitor-bound-cytochrome-bc-1-complexes-for-structure-based-drug-discovery
#8
Kangsa Amporndanai, Rachel M Johnson, Paul M O'Neill, Colin W G Fishwick, Alexander H Jamson, Shaun Rawson, Stephen P Muench, S Samar Hasnain, Svetlana V Antonyuk
Cytochrome bc 1 , a dimeric multi-subunit electron-transport protein embedded in the inner mitochondrial membrane, is a major drug target for the treatment and prevention of malaria and toxoplasmosis. Structural studies of cytochrome bc 1 from mammalian homologues co-crystallized with lead compounds have underpinned structure-based drug design to develop compounds with higher potency and selectivity. However, owing to the limited amount of cytochrome bc 1 that may be available from parasites, all efforts have been focused on homologous cytochrome bc 1 complexes from mammalian species, which has resulted in the failure of some drug candidates owing to toxicity in the host...
March 1, 2018: IUCrJ
https://www.readbyqxmd.com/read/29765598/crystallography-and-the-development-of-therapeutic-medicines
#9
EDITORIAL
Edward N Baker
Advances in crystallography and cryo-EM promise to further enhance the proud role crystallography has played in the development of new medicines.
March 1, 2018: IUCrJ
https://www.readbyqxmd.com/read/29763607/biomedical-applications-of-lumazine-synthase
#10
Yangjie Wei, Prashant Kumar, Newton Wahome, Nicholas J Mantis, C Russell Middaugh
Lumazine synthase (LS) is a family of enzyme involved in the penultimate step in the biosynthesis of riboflavin. Its enzymatic mechanism has been well defined and many LS structures have been solved using x-ray crystallography or cryo-electron microscopy. LS is composed of homo-oligomers, which vary in size and subunit number, including pentamers, decamers, and icosahedral sixty-mers, depending on its species of origin. Research on LS has expanded beyond the initial focus on its enzymatic function to properties related to its oligomeric structure and exceptional conformational stability...
May 12, 2018: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/29761001/multiscale-nonlinear-microscopy-and-widefield-white-light-imaging-enables-rapid-histological-imaging-of-surgical-specimen-margins
#11
Michael G Giacomelli, Tadayuki Yoshitake, Lucas C Cahill, Hilde Vardeh, Liza M Quintana, Beverly E Faulkner-Jones, Jeff Brooker, James L Connolly, James G Fujimoto
The ability to histologically assess surgical specimens in real-time is a long-standing challenge in cancer surgery, including applications such as breast conserving therapy (BCT). Up to 40% of women treated with BCT for breast cancer require a repeat surgery due to postoperative histological findings of close or positive surgical margins using conventional formalin fixed paraffin embedded histology. Imaging technologies such as nonlinear microscopy (NLM), combined with exogenous fluorophores can rapidly provide virtual H&E imaging of surgical specimens without requiring microtome sectioning, facilitating intraoperative assessment of margin status...
May 1, 2018: Biomedical Optics Express
https://www.readbyqxmd.com/read/29760198/cellulose-synthase-stoichiometry-in-aspen-differs-from-arabidopsis-and-norway-spruce
#12
Xueyang Zhang, Pia Guadalupe Dominguez, Manoj Kumar, Joakim Bygdell, Sergey Miroshnichenko, Bjorn Sundberg, Gunnar Wingsle, Totte Niittyla
Cellulose is synthesised at the plasma membrane by cellulose synthase complexes (CSCs) containing cellulose synthases (CESAs). Genetic analysis and CESA isoform quantification indicate that cellulose in the secondary cell walls of Arabidopsis (Arabidopsis thaliana) is synthesised by isoforms CESA4, CESA7 and CESA8 in equimolar amounts. Here, we used quantitative proteomics to investigate whether the CSC model based on Arabidopsis secondary cell wall CESA stoichiometry can be applied to the angiosperm tree aspen (Populus tremula) and the gymnosperm tree Norway spruce (Picea abies)...
May 14, 2018: Plant Physiology
https://www.readbyqxmd.com/read/29758539/bulk-and-dispersed-aqueous-behaviour-of-an-endogenous-lipid-selachyl-alcohol-effect-of-tween-80-and-pluronic-f127-on-nanostructure
#13
Mohammad Younus, Adrian Hawley, Ben J Boyd, Shakila B Rizwan
Tween 80 has been reported to provide a means of targeting drug nanocarriers to the blood- brain barrier. This study investigated the influence of addition of Tween 80 on the formation of different bulk and dispersed lyotropic liquid crystalline phases in selachyl alcohol-based systems. The effect of increasing concentrations of Tween 80 and Pluronic F127 (as a control) (0-25% w/w relative to SA) on the bulk phase behaviour and dispersions of selachyl alcohol (SA) were investigated using small angle X-ray scattering, dynamic light scattering, and cryogenic transmission electron microscopy...
May 7, 2018: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29758301/lipid-roles-in-herg-function-and-interactions-with-drugs
#14
REVIEW
Meruyert Kudaibergenova, Laura L Perissinotti, Sergei Y Noskov
Human-ether-a-go-go-related channel (hERG) is a voltage gated potassium channel (Kv 11.1) abundantly expressed in heart and brain tissues. In addition to playing an important role in mediation of repolarizing K+ currents (IKr ) in Action Potential (AP), hERG is notorious for its propensity to interact with various medications. The drug-induced block of K+ currents across hERG channel are strongly associated with dysrhythmic conditions collectively known as drug-induced long-QT-syndrome. The recent availability of the high-resolution Cryo-EM structures for the hERG channel has provided unique opportunity to resolve structural mechanisms involved into the process of voltage-gating of hERG channels, map various roles played by components of ventricular and neuronal membranes and then to connect it to cellular pathways through which diverse chemical compounds might be affecting function of the channel...
May 11, 2018: Neuroscience Letters
https://www.readbyqxmd.com/read/29757285/microfluidic-chips-for-in-situ-crystal-x-ray-diffraction-and-in-situ-dynamic-light-scattering-for-serial-crystallography
#15
Yannig Gicquel, Robin Schubert, Svetlana Kapis, Gleb Bourenkov, Thomas Schneider, Markus Perbandt, Christian Betzel, Henry N Chapman, Michael Heymann
This protocol describes fabricating microfluidic devices with low X-ray background optimized for goniometer based fixed target serial crystallography. The devices are patterned from epoxy glue using soft lithography and are suitable for in situ X-ray diffraction experiments at room temperature. The sample wells are lidded on both sides with polymeric polyimide foil windows that allow diffraction data collection with low X-ray background. This fabrication method is undemanding and inexpensive. After the sourcing of a SU-8 master wafer, all fabrication can be completed outside of a cleanroom in a typical research lab environment...
April 24, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29755750/ionic-scattering-factors-of-atoms-that-compose-biological-molecules
#16
Koji Yonekura, Rei Matsuoka, Yoshiki Yamashita, Tsutomu Yamane, Mitsunori Ikeguchi, Akinori Kidera, Saori Maki-Yonekura
Ionic scattering factors of atoms that compose biological molecules have been computed by the multi-configuration Dirac-Fock method. These ions are chemically unstable and their scattering factors had not been reported except for O- . Yet these factors are required for the estimation of partial charges in protein molecules and nucleic acids. The electron scattering factors of these ions are particularly important as the electron scattering curves vary considerably between neutral and charged atoms in the spatial-resolution range explored in structural biology...
May 1, 2018: IUCrJ
https://www.readbyqxmd.com/read/29754826/atomic-resolution-cryo-em-structure-of-%C3%AE-galactosidase
#17
Alberto Bartesaghi, Cecilia Aguerrebere, Veronica Falconieri, Soojay Banerjee, Lesley A Earl, Xing Zhu, Nikolaus Grigorieff, Jacqueline L S Milne, Guillermo Sapiro, Xiongwu Wu, Sriram Subramaniam
The advent of direct electron detectors has enabled the routine use of single-particle cryo-electron microscopy (EM) approaches to determine structures of a variety of protein complexes at near-atomic resolution. Here, we report the development of methods to account for local variations in defocus and beam-induced drift, and the implementation of a data-driven dose compensation scheme that significantly improves the extraction of high-resolution information recorded during exposure of the specimen to the electron beam...
April 30, 2018: Structure
https://www.readbyqxmd.com/read/29754813/structure-of-an-ancient-respiratory-system
#18
Hongjun Yu, Chang-Hao Wu, Gerrit J Schut, Dominik K Haja, Gongpu Zhao, John W Peters, Michael W W Adams, Huilin Li
Hydrogen gas-evolving membrane-bound hydrogenase (MBH) and quinone-reducing complex I are homologous respiratory complexes with a common ancestor, but a structural basis for their evolutionary relationship is lacking. Here, we report the cryo-EM structure of a 14-subunit MBH from the hyperthermophile Pyrococcus furiosus. MBH contains a membrane-anchored hydrogenase module that is highly similar structurally to the quinone-binding Q-module of complex I while its membrane-embedded ion-translocation module can be divided into a H+ - and a Na+ -translocating unit...
April 25, 2018: Cell
https://www.readbyqxmd.com/read/29754486/nanoparticle-wettability-influences-nanoparticle-phospholipid-interactions
#19
Nagarjun Konduru, Flavia Damiani, Svetla Stoilova-McPhie, Jason Tresback, Georgios Pyrgiotakis, Thomas C Donaghey, Philip Demokritou, Joseph Brain, Ramon M Molina
We explored the influence of nanoparticle (NP) surface charge and hydrophobicity on nanoparticle-biomolecule interactions by measuring the composition of adsorbed phospholipids on four NPs, namely positively charged CeO2 and ZnO, and negatively charged BaSO4 and silica-coated CeO2 after exposure to bronchoalveolar lavage fluid (BALf) obtained from rats, and to a mixture of neutral dipalmitoyl phosphatidylcholine (DPPC) and negatively charged dipalmitoyl phosphatidic acid (DPPA). The resulting NP-lipid interactions were examined by cryogenic transmission electron microscopy (cryo-TEM) and by atomic force microscopy (AFM)...
May 12, 2018: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/29754178/ev-microvesicles-micrornas-and-stem-cells-in-cancer
#20
Jacob A Tickner, Derek J Richard, Kenneth J O'Byrne
The role of extracellular vesicles (EV) in carcinogenesis has become the focus of much research. These microscopic messengers have been found to regulate immune system function, particularly in tumorigenesis, as well as conditioning future metastatic sites for the attachment and growth of tumor tissue. Through an interaction with a range of host tissues, EVs are able to generate a pro-tumor environment that is essential for tumorigenesis. These small nanovesicles are an ideal candidate for a non-invasive indicator of pathogenesis and/or disease progression as they can display individualized nucleic acid, protein, and lipid expression profiles that are often reflective of disease state, and can be easily detected in bodily fluids, even after extended cryo-storage...
2018: Advances in Experimental Medicine and Biology
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