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Journal of Structural Biology

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https://www.readbyqxmd.com/read/29630922/delivery-of-femtolitre-droplets-using-surface-acoustic-wave-based-atomisation-for-cryo-em-grid-preparation
#1
Dariush Ashtiani, Hari Venugopal, Matthew Belousoff, Bradley Spicer, Johnson Mak, Adrian Neild, Alex de Marco
Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required...
April 6, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29626512/substrate-bound-structures-of-a-ketoreductase-from-amphotericin-modular-polyketide-synthase
#2
Chenguang Liu, Meijuan Yuan, Xu Xu, Lei Wang, Adrian T Keatinge-Clay, Zixin Deng, Shuangjun Lin, Jianting Zheng
Ketoreductase (KR) domains of modular polyketide synthases (PKSs) control the stereochemistry of C2 methyl and C3 hydroxyl substituents of polyketide intermediates. To understand the molecular basis of stereocontrol exerted by KRs, the crystal structure of a KR from the second module of the amphotericin PKS (AmpKR2) complexed with NADP+ and 2-methyl-3-oxopentanoyl-pantetheine was solved. This first ternary structure provides direct evidence to the hypothesis that a substrate enters into the active site of an A-type KR from the side opposite the coenzyme to generate an L-hydroxyl substituent...
April 4, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29605571/structural-and-functional-analysis-of-erwinia-amylovora-srld-the-first-crystal-structure-of-a-sorbitol-6-phosphate-2-dehydrogenase
#3
Marco Salomone-Stagni, Joseph D Bartho, Eeshan Kalita, Martin Rejzek, Robert A Field, Dom Bellini, Martin A Walsh, Stefano Benini
Sorbitol-6-phosphate 2-dehydrogenases (S6PDH) catalyze the interconversion of d-sorbitol 6-phosphate to d-fructose 6-phosphate. In the plant pathogen Erwinia amylovora the S6PDH SrlD is used by the bacterium to utilize sorbitol, which is used for carbohydrate transport in the host plants belonging to the Amygdaloideae subfamily (e.g., apple, pear, and quince). We have determined the crystal structure of S6PDH SrlD at 1.84 Å resolution, which is the first structure of an EC 1.1.1.140 enzyme. Kinetic data show that SrlD is much faster at oxidizing d-sorbitol 6-phosphate than in reducing d-fructose 6-phosphate, however, equilibrium analysis revealed that only part of the d-sorbitol 6-phosphate present in the in vitro environment is converted into d-fructose 6-phosphate...
March 29, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29605570/a-new-hiv-1-rev-structure-optimizes-interaction-with-target-rna-rre-for-nuclear-export
#4
Norman R Watts, Elif Eren, Xiaolei Zhuang, Yun-Xing Wang, Alasdair C Steven, Paul T Wingfield
HIV-1 Rev mediates the nuclear export of unspliced and partially-spliced viral transcripts for the production of progeny genomes and structural proteins. In this process, four (or more) copies of Rev assemble onto a highly-structured 351-nt region in such viral transcripts, theRevresponseelement (RRE). How this occurs is not known. The Rev assembly domain has a helical-hairpin structure which associates through three (A-A, B-B and C-C) interfaces. The RRE has the topology of an upper-case letter A, with the two known Rev binding sites mapping onto the legs of the A...
March 29, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29604451/amyloid-structure-of-high-order-assembly-of-leucine-rich-amelogenin-revealed-by-solid-state-nmr
#5
Cheng-Wei Ma, Jing Zhang, Xing-Qi Dong, Jun-Xia Lu
High-order assemblies of amelogenin, the major protein in enamel protein matrix, are believed to act as the template for enamel mineral formation. The Leucine-rich amelogenin (LRAP) is a natural splice-variant of amelogenin, a functional protein in vivo, containing conserved domains of amelogenin. In this work, we showed LRAP aggregates hierarchically into assemblies with various sizes including scattered beads, beads-on-a-string and gel-like precipitations in the presence of both calcium and phosphate ions...
March 28, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29601957/structure-and-electrochemistry-of-proteins-harboring-iron-sulfur-clusters-of-different-nuclearities-part-iii-4fe-4s-3fe-4s-and-2fe-2s-iron-sulfur-proteins
#6
REVIEW
Piero Zanello
A systematic rationalization of the hundreds of proteins harboring iron-sulfur clusters and able to exhibit the most diverse biological functions is missing. In this picture we have already reviewed structure/electrochemistry of metalloproteins expressing single types of iron-sulfur centres [namely, {Fe(Cys)4 }, { [Fe2 S2 ](Cys)4 }, {[Fe2 S2 ](Cys)3 (X)} (X = Asp, Arg, His), [Fe2 S2 ](Cys)2 (His)2 }, { [Fe3 S4 ](Cys)3 }, { [Fe4 S4 ](Cys)4 } and { [Fe4 S4 ](Sγ Cys )3 (nonthiolate ligand)}] and their synthetic analogs...
March 27, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29551714/blind-estimation-of-ded-camera-gain-in-electron-microscopy
#7
C O S Sorzano, E Fernández-Giménez, V Peredo-Robinson, J Vargas, T Majtner, G Caffarena, J Otón, J L Vilas, J M de la Rosa-Trevín, R Melero, J Gómez-Blanco, J Cuenca, L Del Cano, P Conesa, R Marabini, J M Carazo
The introduction of Direct Electron Detector (DED) videos in the Electron Microscope field has boosted Single Particle Analysis to a point in which it is currently considered to be a key technique in Structural Biology. In this article we introduce an approach to estimate the DED camera gain at each pixel from the movies themselves. This gain is needed to have the set of recorded frames into a coherent gray level range, homogeneous over the whole image. The algorithm does not need any other input than the DED movie itself, being capable of providing an estimate of the camera gain image, helping to identify dead pixels and cases of incorrectly calibrated cameras...
March 15, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29551713/improved-sample-dispersion-in-cryo-em-using-perpetually-hydrated-graphene-oxide-flakes
#8
Martin Cheung, Hidehito Adaniya, Cathal Cassidy, Masao Yamashita, Kun-Lung Li, Seita Taba, Tsumoru Shintake
For many macromolecular complexes, the inability to uniformly disperse solubilized specimen particles within vitreous ice films precludes their analysis by cryo-electron microscopy (cryo-EM). Here, we introduce a sample preparation process using "perpetually-hydrated" graphene oxide flakes as particle support films, and report vastly improved specimen dispersion. Furthermore, we provide evidence that the presence of graphene oxide flakes in vitreous ice results in a significant reduction in electron beam-induced specimen decomposition...
March 15, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29549035/structure-of-a-prokaryotic-sefir-domain-reveals-two-novel-sefir-sefir-interaction-modes
#9
Hui Yang, Yun Zhu, Xing Chen, Xiaoxia Li, Sheng Ye, Rongguang Zhang
SEFIR domain-containing proteins are crucial for mammalian adaptive immunity. As a unique intracellular signaling domain, the SEFIR-SEFIR interactions mediate physical protein-protein interactions in the immune signaling network, especially the IL-17- and IL-25-mediated pathways. However, due to the lack of structural information, the detailed molecular mechanism for SEFIR-SEFIR assembly remains unclear. In the present study, we solved the crystal structures of a prokaryotic SEFIR domain from Bacillus cereus F65185 (BcSEFIR), which SEFIR domain located at the N terminusof protein...
March 13, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29545204/baculovirus-driven-protein-expression-in-insect-cells-a-benchmarking-study
#10
Peggy Stolt-Bergner, Christian Benda, Tim Bergbrede, Hüseyin Besir, Patrick H N Celie, Cindy Chang, David Drechsel, Ariane Fischer, Arie Geerlof, Barbara Giabbai, Joop van den Heuvel, Georg Huber, Wolfgang Knecht, Anita Lehner, Regis Lemaitre, Kristina Nordén, Gwynn Pardee, Ines Racke, Kim Remans, Astrid Sander, Judith Scholz, Magda Stadnik, Paola Storici, Daniel Weinbruch, Isabel Zaror, Linda H L Lua, Sabine Suppmann
Baculovirus-insect cell expression system has become one of the most widely used eukaryotic expression systems for heterologous protein production in many laboratories. The availability of robust insect cell lines, serum-free media, a range of vectors and commercially-packaged kits have supported the demand for maximizing the exploitation of the baculovirus-insect cell expression system. Naturally, this resulted in varied strategies adopted by different laboratories to optimize protein production. Most laboratories have preference in using either the E...
March 12, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29526782/solution-structure-of-an-archaeal-duf61-family-protein-sso0941-encoded-by-a-gene-in-the-operon-of-box-c-d-rna-protein-complexes
#11
Tao Zhou, Xingzhe Yao, Jinfeng Wang, Yingang Feng
Domain of unknown function 61 (DUF61) family proteins widely exist in archaea and the genes of DUF61 proteins in crenarchaea are in an operon containing two genes of box C/D RNA protein complexes. Here we report the solution NMR structure of DUF61 family member protein SSO0941, from the hyperthermophilic archaeon Sulfolobus solfataricus. SSO0941 has a rigid core structure and flexible N- and C-terminal regions as well as a negatively-charged independent C-terminal helix. The core structure consists of N- and C-terminal subdomains, in which the C-terminal subdomain shows significant structural similarity with several nucleic acid binding proteins...
March 8, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29522817/gaussian-input-gaussian-mixture-model-for-representing-density-maps-and-atomic-models
#12
Takeshi Kawabata
A new Gaussian mixture model (GMM) has been developed for better representations of both atomic models and electron microscopy 3D density maps. The standard GMM algorithm employs an EM algorithm to determine the parameters. It accepted a set of 3D points with weights, corresponding to voxel or atomic centers. Although the standard algorithm worked reasonably well; however, it had three problems. First, it ignored the size (voxel width or atomic radius) of the input, and thus it could lead to a GMM with a smaller spread than the input...
March 6, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29505829/structural-basis-for-partition-of-the-cyclodipeptide-synthases-into-two-subfamilies
#13
Gabrielle Bourgeois, Jérôme Seguin, Morgan Babin, Pascal Belin, Mireille Moutiez, Yves Mechulam, Muriel Gondry, Emmanuelle Schmitt
Cyclodipeptide synthases (CDPSs) use two aminoacyl-tRNAs to catalyze the formation of two peptide bonds leading to cyclodipeptides that can be further used for the synthesis of diketopiperazines. It was shown that CDPSs fall into two subfamilies, NYH and XYP, characterized by the presence of specific sequence signatures. However, current understanding of CDPSs only comes from studies of enzymes from the NYH subfamily. The present study reveals the crystal structures of three CDPSs from the XYP subfamily. Comparison of the XYP and NYH enzymes shows that the two subfamilies mainly differ in the first half of their Rossmann fold...
March 2, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29501724/%C3%AE-cgrp-another-amyloidogenic-member-of-the-cgrp-family
#14
Paraskevi L Tsiolaki, Georgia I Nasi, Fotis A Baltoumas, Nikolaos N Louros, Vassiliki Magafa, Stavros J Hamodrakas, Vassiliki A Iconomidou
The Calcitonin-gene related peptide (CGRP) family is a group of peptide hormones, which consists of IAPP, calcitonin, adrenomedullin, intermedin, αCGRP and βCGRP. IAPP and calcitonin have been extensively associated with the formation of amyloid fibrils, causing Type 2 Diabetes and Medullary Thyroid Carcinoma, respectively. In contrast, the potential amyloidogenic properties of αCGRP still remain unexplored, although experimental trials have indicated its presence in deposits, associated with the aforementioned disorders...
March 1, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29501472/interplay-between-membrane-curvature-and-protein-conformational-equilibrium-investigated-by-solid-state-nmr
#15
Shu Y Liao, Myungwoon Lee, Mei Hong
Many membrane proteins sense and induce membrane curvature for function, but structural information about how proteins modulate their structures to cause membrane curvature is sparse. We review our recent solid-state NMR studies of two virus membrane proteins whose conformational equilibrium is tightly coupled to membrane curvature. The influenza M2 proton channel has a drug-binding site in the transmembrane (TM) pore. Previous chemical shift data indicated that this pore-binding site is lost in an M2 construct that contains the TM domain and a curvature-inducing amphipathic helix...
February 28, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29486249/exploring-applications-of-crowdsourcing-to-cryo-em
#16
Jacob Bruggemann, Gabriel C Lander, Andrew I Su
Extraction of particles from cryo-electron microscopy (cryo-EM) micrographs is a crucial step in processing single-particle datasets. Although algorithms have been developed for automatic particle picking, these algorithms generally rely on two-dimensional templates for particle identification, which may exhibit biases that can propagate artifacts through the reconstruction pipeline. Manual picking is viewed as a gold-standard solution for particle selection, but it is too time-consuming to perform on data sets of thousands of images...
February 24, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29481850/coiled-coil-oligomerization-controls-nanodomain-organization-of-the-plasma-membrane-remorins
#17
Denis Martinez, Anthony Legrand, Julien Gronnier, Marion Decossas, Paul Gouguet, Olivier Lambert, Mélanie Berbon, Loris Verron, Axelle Grélard, Veronique Germain, Antoine Loquet, Sébastien Mongrand, Birgit Habenstein
REMORINs are nanodomain-organized proteins located in the plasma membrane and involved in cellular responses in plants. The dynamic assembly of the membrane nanodomains represents an essential tool of the versatile membrane barriers to control and modulate cellular functions. Nevertheless, the assembly mechanisms and protein organization strategies of nanodomains are poorly understood and many structural aspects are difficult to visualize. Using an ensemble of biophysical approaches, including solid-state nuclear magnetic resonance, cryo-electron microscopy and in vivo confocal imaging, we provide first insights on the role and the structural mechanisms of REMORIN trimerization...
February 23, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29477758/an-automated-workflow-for-segmenting-single-adult-cardiac-cells-from-large-volume-serial-block-face-scanning-electron-microscopy-data
#18
Akter Hussain, Shouryadipta Ghosh, Siavash Beikoghli Kalkhoran, Derek J Hausenloy, Eric Hanssen, Vijay Rajagopal
This paper presents a new algorithm to automatically segment the myofibrils, mitochondria and nuclei within single adult cardiac cells that are part of a large serial-block-face scanning electron microscopy (SBF-SEM) dataset. The algorithm only requires a set of manually drawn contours that roughly demarcate the cell boundary at routine slice intervals (every 50th , for example). The algorithm correctly classified pixels within the single cell with 97% accuracy when compared to manual segmentations. One entire cell and the partial volumes of two cells were segmented...
February 22, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29454111/combining-rosetta-with-molecular-dynamics-md-a-benchmark-of-the-md-based-ensemble-protein-design
#19
Jan Ludwiczak, Adam Jarmula, Stanislaw Dunin-Horkawicz
Computational protein design is a set of procedures for computing amino acid sequences that will fold into a specified structure. Rosetta Design, a commonly used software for protein design, allows for the effective identification of sequences compatible with a given backbone structure, while molecular dynamics (MD) simulations can thoroughly sample near-native conformations. We benchmarked a procedure in which Rosetta design is started on MD-derived structural ensembles and showed that such a combined approach generates 20-30% more diverse sequences than currently available methods with only a slight increase in computation time...
February 14, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29428558/structure-and-electrochemistry-of-proteins-harboring-iron-sulfur-clusters-of-different-nuclearities-part-ii-4fe-4s-and-3fe-4s-iron-sulfur-proteins
#20
REVIEW
Piero Zanello
In the context of the plethora of proteins harboring iron-sulfur clusters we have already reviewed structure/electrochemistry of metalloproteins expressing single types of iron-sulfur clusters (namely: {Fe(Cys)4}, { [Fe2S2](Cys)4}, { [Fe2S2](Cys)3(X)} (X = Asp, Arg, His), { [Fe2S2](Cys)2(His)2}, { [Fe3S4](Cys)3}, { [Fe4S4](Cys)4} and { [Fe4S4](SγCys)3(nonthiolate ligand) } cores) and their synthetic analogs. More recently we are focussing on structure/electrochemistry of metalloproteins harboring iron-sulfur centres of different nuclearities...
February 8, 2018: Journal of Structural Biology
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