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https://www.readbyqxmd.com/read/29046050/immunofluorescent-characterization-of-non-myelinating-schwann-cells-and-their-interactions-with-immune-cells-in-mouse-mesenteric-lymph-node
#1
Zhongli Shi, Wayne K Greene, Philip K Nicholls, Dailun Hu, Janina E E Tirnitz-Parker, Qionglan Yuan, Changfu Yin, Bin Ma
The central nervous system (CNS) influences the immune system in a general fashion by regulating the systemic concentration of humoral substances, whereas the autonomic nervous system communicates specifically with the immune system according to local interactions. Data concerning the mechanisms of this bidirectional crosstalk of the peripheral nervous system (PNS) and immune system remain limited. To gain a better understanding of local interactions of the PNS and immune system, we have used immunofluorescent staining of glial fibrillary acidic protein (GFAP), coupled with confocal microscopy, to investigate the non-myelinating Schwann cell (NMSC)-immune cell interactions in mouse mesenteric lymph nodes...
August 7, 2017: European Journal of Histochemistry: EJH
https://www.readbyqxmd.com/read/29045929/deciphering-the-complexation-process-of-a-fluoroquinolone-antibiotic-levofloxacin-with-bovine-serum-albumin-in-the-presence-of-additives
#2
Amandeep Kaur, Imran Ahmd Khan, Parampaul Kaur Banipal, Tarlok Singh Banipal
The current work aims to explore the thermodynamic and conformational aspects for the binding of fluoroquinolone antibacterial drug, levofloxacin (LFC), with bovine serum albumin (BSA) using calorimetric, spectroscopic (UV-visible, fluorescence, circular dichroism, and (1)H NMR), dynamic light scattering (DLS) and computational methods (molecular docking). The binding of LFC with BSA at two sequential sites with higher affinity (~10(3)M(-1)) at the first site has been explored by calorimetry whereas the binding at a single site with affinity of the order of ~10(4)M(-1) has been observed from fluorescence spectroscopy...
October 9, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/29045898/peptidoglycan-sensing-receptors-trigger-the-formation-of-functional-amyloids-of-the-adaptor-protein-imd-to-initiate-drosophila-nf-%C3%AE%C2%BAb-signaling
#3
Anni Kleino, Nancy F Ramia, Gunes Bozkurt, Yanfang Shen, Himani Nailwal, Jing Huang, Johanna Napetschnig, Monique Gangloff, Francis Ka-Ming Chan, Hao Wu, Jixi Li, Neal Silverman
In the Drosophila immune response, bacterial derived diaminopimelic acid-type peptidoglycan binds the receptors PGRP-LC and PGRP-LE, which through interaction with the adaptor protein Imd leads to activation of the NF-κB homolog Relish and robust antimicrobial peptide gene expression. PGRP-LC, PGRP-LE, and Imd each contain a motif with some resemblance to the RIP Homotypic Interaction Motif (RHIM), a domain found in mammalian RIPK proteins forming functional amyloids during necroptosis. Here we found that despite sequence divergence, these Drosophila cryptic RHIMs formed amyloid fibrils in vitro and in cells...
October 17, 2017: Immunity
https://www.readbyqxmd.com/read/29045874/dynamic-scaling-analysis-of-molecular-motion-within-the-lat-grb2-sos-protein-network-on%C3%A2-membranes
#4
William Y C Huang, Han-Kuei Chiang, Jay T Groves
Biochemical signaling pathways often involve proteins with multiple, modular interaction domains. Signaling activates binding sites, such as by tyrosine phosphorylation, which enables protein recruitment and growth of networked protein assemblies. Although widely observed, the physical properties of the assemblies, as well as the mechanisms by which they function, remain largely unknown. Here we examine molecular mobility within LAT:Grb2:SOS assemblies on supported membranes by single-molecule tracking. Trajectory analysis reveals a discrete temporal transition to subdiffusive motion below a characteristic timescale, indicating that the LAT:Grb2:SOS assembly has the dynamical structure of a loosely entangled polymer...
October 17, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/29045865/cooperative-nucleotide-binding-in-hsp90-and-its-regulation-by-aha1
#5
Philipp Wortmann, Markus Götz, Thorsten Hugel
The function of the molecular chaperone Hsp90 depends on large conformational changes, the rearrangement of local motifs, and the binding and hydrolysis of ATP. The size and complexity of the Hsp90 system impedes the detailed investigation of their interplay using standard methods. To overcome this limitation, we developed a three-color single-molecule FRET assay to study the interaction of Hsp90 with a fluorescently labeled reporter nucleotide in detail. It allows us to directly observe the cooperativity between the two nucleotide binding pockets in the protein dimer...
October 17, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/29045864/molecular-simulations-suggest-a-force-dependent-mechanism-of-vinculin-activation
#6
Li Sun, Jeffrey K Noel, Herbert Levine, José N Onuchic
Focal adhesions are dynamic constructs at the leading edge of migrating cells, linking them to the extracellular matrix and enabling force sensing and transmission. The lifecycle of a focal adhesion is a highly coordinated process involving spatial and temporal variations of protein composition, interaction, and cellular tension. The assembly of focal adhesions requires the recruitment and activation of vinculin. Vinculin is present in the cytoplasm in an autoinhibited conformation in which its tail is held pincerlike by its head domains, further stabilized by two high-affinity head-tail interfaces...
October 17, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/29045862/the-two-state-prehensile-tail-of-the-antibacterial-toxin-colicin-n
#7
Christopher L Johnson, Alexandra S Solovyova, Olli Hecht, Colin Macdonald, Helen Waller, J Günter Grossmann, Geoffrey R Moore, Jeremy H Lakey
Intrinsically disordered regions within proteins are critical elements in many biomolecular interactions and signaling pathways. Antibacterial toxins of the colicin family, which could provide new antibiotic functions against resistant bacteria, contain disordered N-terminal translocation domains (T-domains) that are essential for receptor binding and the penetration of the Escherichia coli outer membrane. Here we investigate the conformational behavior of the T-domain of colicin N (ColN-T) to understand why such domains are widespread in toxins that target Gram-negative bacteria...
October 17, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/29045861/effects-of-nanopore-charge-decorations-on-the-translocation-dynamics-of-dna
#8
Ining Jou, Murugappan Muthukumar
We have investigated the dynamics of single-stranded DNA as it translocates through charge-mutated protein nanopores. Translocation of DNA is a crucial step in nanopore-based sequencing platforms, where control over translocation speed remains one of the main challenges. Taking advantage of the interactions between negatively charged DNA and positively charged amino acid residues, the translocation speed of DNA can be manipulated by deliberate charge decorations inside the nanopore. We employed coarse-grained Langevin dynamics simulations to monitor the step-by-step movement of DNA through different mutations of α-hemolysin protein nanopores...
October 17, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/29045859/role-of-condensing-particles-in-polymer-confinement-a-model-for-virus-packed-minichromosomes
#9
Sanjin Marion, Carmen San Martín, Antonio Šiber
Confined mixtures of a polymer and nonspecifically binding particles (condensers) are studied as models for viruses containing double-stranded DNA (polymer) and condensing proteins (particles). We explore a model in which all interactions between the packed content (polymer and particles) and its confinement are purely repulsive, with only a short-range attraction between the condensers and polymer to simulate binding. In the range of physical parameters applicable to viruses, the model predicts reduction of pressure in the system effected by the condensers, despite the reduction in free volume...
October 17, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/29045836/arc-requires-psd95-for-assembly-into-postsynaptic-complexes-involved-with-neural-dysfunction-and-intelligence
#10
Esperanza Fernández, Mark O Collins, René A W Frank, Fei Zhu, Maksym V Kopanitsa, Jess Nithianantharajah, Sarah A Lemprière, David Fricker, Kathryn A Elsegood, Catherine L McLaughlin, Mike D R Croning, Colin Mclean, J Douglas Armstrong, W David Hill, Ian J Deary, Giulia Cencelli, Claudia Bagni, Menachem Fromer, Shaun M Purcell, Andrew J Pocklington, Jyoti S Choudhary, Noboru H Komiyama, Seth G N Grant
Arc is an activity-regulated neuronal protein, but little is known about its interactions, assembly into multiprotein complexes, and role in human disease and cognition. We applied an integrated proteomic and genetic strategy by targeting a tandem affinity purification (TAP) tag and Venus fluorescent protein into the endogenous Arc gene in mice. This allowed biochemical and proteomic characterization of native complexes in wild-type and knockout mice. We identified many Arc-interacting proteins, of which PSD95 was the most abundant...
October 17, 2017: Cell Reports
https://www.readbyqxmd.com/read/29045822/individual-effects-of-the-copia-and-gypsy-enhancer-and-insulator-on-chromatin-marks-erna-synthesis-and-binding-of-insulator-proteins-in-transfected-genetic-constructs
#11
Daria M Fedoseeva, Olga V Kretova, Maria A Gorbacheva, Nickolai A Tchurikov
Enhancers and insulators are involved in the regulation of gene expression, but the basic underlying mechanisms of action of these elements are unknown. We analyzed the individual effects of the enhancer and the insulator from Drosophila mobile elements copia [enh(copia)] and gypsy using transfected genetic constructs in S2 cells. This system excludes the influence of genomic cis regulatory elements. The enhancer-induced synthesis of 350-1050-nt-long enhancer RNAs (eRNAs) and H3K4me3 and H3K18ac marks, mainly in the region located about 300bp downstream of the enhancer...
October 15, 2017: Gene
https://www.readbyqxmd.com/read/29045787/mechanistic-origin-of-the-combined-effect-of-surfaces-and-mechanical-agitation-on-amyloid-formation
#12
Fulvio Grigolato, Claudio Colombo, Raffaele Ferrari, Lenka Rezabkova, Paolo Arosio
Interactions between proteins and surfaces in combination with hydrodynamic flow and mechanical agitation can often trigger the conversion of soluble peptides and proteins into aggregates, including amyloid fibrils. Despite the extensive literature on the empirical effects of surfaces and mechanical forces on the formation of amyloids, the molecular details of the mechanisms underlying this behavior are still elusive. This limitation is, in part, due to the complex reaction network underlying the formation of amyloids, where several microscopic reactions of nucleation and growth can occur both at the interfaces and in bulk...
October 18, 2017: ACS Nano
https://www.readbyqxmd.com/read/29045780/mechanisms-of-activation-and-subunit-release-in-ca2-calmodulin-dependent-protein-kinase-ii
#13
Filippo Pullara, Eliana Karina Asciutto, Ignacio Jose General
Calcium/Calmodulin dependent protein kinase II is an enzyme involved in many different functions, including the so-called long-term potentiation, a mechanism that strengthens synapses in a persistent mode, and is believed to be a basic cellular mechanism for memory formation. Here we study the conformational changes of the enzyme due to phosphorylation of some key residues that are believed to drive the transition from an inhibited to an active state; it is this active state the one associated with long-term potentiation...
October 18, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/29045748/conserved-asymmetry-underpins-homodimerization-of-dicer-associated-double-stranded-rna-binding-proteins
#14
Alex Heyam, Claire E Coupland, Clément Dégut, Ruth A Haley, Nicola J Baxter, Leonhard Jakob, Pedro M Aguiar, Gunter Meister, Michael P Williamson, Dimitris Lagos, Michael J Plevin
Double-stranded RNA-binding domains (dsRBDs) are commonly found in modular proteins that interact with RNA. Two varieties of dsRBD exist: canonical Type A dsRBDs interact with dsRNA, while non-canonical Type B dsRBDs lack RNA-binding residues and instead interact with other proteins. In higher eukaryotes, the microRNA biogenesis enzyme Dicer forms a 1:1 association with a dsRNA-binding protein (dsRBP). Human Dicer associates with HIV TAR RNA-binding protein (TRBP) or protein activator of PKR (PACT), while Drosophila Dicer-1 associates with Loquacious (Loqs)...
October 17, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29045742/white-leaf-and-panicle-2-encoding-a-pep-associated-protein-is-required-for-chloroplast-biogenesis-under-heat-stress-in-rice
#15
Yusong Lv, Gaoneng Shao, Jiehua Qiu, Guiai Jiao, Zhonghua Sheng, Lihong Xie, Yawen Wu, Shaoqing Tang, Xiangjin Wei, Peisong Hu, Robert Sharwood
The plastid-encoded RNA polymerase (PEP) plays an important role in the transcription machinery of mature chloroplasts, yet details of its function remain elusive in rice. Here, we identified a novel PEP-associated protein (PAP), WLP2, based on its two allelic white leaf and panicle mutants, wlp2s and wlp2w. The two mutants were albino lethal at high temperatures and showed decreased chlorophyll accumulation, abnormal chloroplast ultrastructure, and attenuated photosynthetic activity. Map-based cloning suggested that WLP2 encodes a putative pfkB-type carbohydrate kinase family protein, which is homologous to fructokinase-like 1 (AtFLN1) in Arabidopsis...
October 16, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/29045730/ribosomal-protein-ntrpl17-interacts-with-kinesin-12-family-protein-ntkrp-and-functions-in-the-regulation-of-embryo-seed-size-and-radicle-growth
#16
Shujuan Tian, Jingjing Wu, Yuan Liu, Xiaorong Huang, Fen Li, Zhaodan Wang, Meng-Xiang Sun, Miriam Gifford
We previously reported that a novel motor protein belonging to the kinesin-12 family, NtKRP, displays critical roles in regulating embryo and seed size establishment. However, it remains unknown exactly how NtKRP contributes to this developmental process. Here, we report that a 60S ribosomal protein NtRPL17 directly interacts with NtKRP. The phenotypes of NtRPL17 RNAi lines show notable embryo and seed size reduction. Structural observations of the NtRPL17-silenced embryos/seeds reveal that the embryo size reduction is due to a decrease in cell number...
October 14, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/29045717/interaction-between-calcium-and-potassium-modulates-elongation-rate-in-cotton-fiber-cells
#17
Kai Guo, Lili Tu, Yonghui He, Jinwu Deng, Maojun Wang, Hui Huang, Zhonghua Li, Xianlong Zhang, Miriam Gifford
Calcium (Ca2+) is necessary for fiber cell development in cotton (Gossypium hirsutum), both as a cell wall structural component and for environmental signaling responses. It is also known that potassium (K+) plays a critical role in cotton fiber cell elongation. However, it is unclear whether Ca2+ integrates its activities with K+ to regulate fiber elongation. Here, we report the novel discovery that Ca2+ deficiency, when integrated with K+ signaling, promotes fiber elongation. Using inductively coupled plasma-mass spectrometry (ICP-MS), we determined dynamic profiles of the ionome in ovules and fibers at different developmental stages, and found that a high accumulation of macro-elements, but not Ca2+, was associated with longer fibers...
October 13, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/29045568/loss-of-protein-kinase-novel-1-pkn1-is-associated-with-mild-systolic-and-diastolic-contractile-dysfunction-increased-phospholamban-thr17-phosphorylation-and-exacerbated-ischaemia-reperfusion-injury
#18
Asvi A Francois, Kofo Obasanjo-Blackshire, James E Clark, Andrii Boguslavskyi, Mark R Holt, Peter Parker, Michael S Marber, Richard J Heads
Aims: PKN1 is a stress-responsive protein kinase acting downstream of small GTP-binding proteins of the Rho/Rac family. The aim was to determine its role in endogenous cardioprotection. Methods and Results: Hearts from PKN1 knockout (KO) or wild type (WT) littermate control mice were perfused in Langendorff mode and subjected to global ischemia and reperfusion (I/R). Myocardial infarct size was doubled in PKN1 KO hearts compared to WT hearts. PKN1 was basally phosphorylated on the activation loop Thr778 PDK1 target site which was unchanged during I/R...
October 16, 2017: Cardiovascular Research
https://www.readbyqxmd.com/read/29045470/structural-and-functional-analysis-of-smo-1-the-sumo-homolog-in-caenorhabditis-elegans
#19
Parag Surana, Chandrakala M Gowda, Vasvi Tripathi, Limor Broday, Ranabir Das
SUMO proteins are important post-translational modifiers involved in multiple cellular pathways in eukaryotes, especially during the different developmental stages in multicellular organisms. The nematode C. elegans is a well known model system for studying metazoan development and has a single SUMO homolog, SMO-1. Interestingly, SMO-1 modification is linked to embryogenesis and development in the nematode. However, high-resolution information about SMO-1 and the mechanism of its conjugation is lacking. In this work, we report the high-resolution three dimensional structure of SMO-1 solved by NMR spectroscopy...
2017: PloS One
https://www.readbyqxmd.com/read/29045465/identifying-protein-complex-by-integrating-characteristic-of-core-attachment-into-dynamic-ppi-network
#20
Xianjun Shen, Li Yi, Xingpeng Jiang, Tingting He, Jincai Yang, Wei Xie, Po Hu, Xiaohua Hu
How to identify protein complex is an important and challenging task in proteomics. It would make great contribution to our knowledge of molecular mechanism in cell life activities. However, the inherent organization and dynamic characteristic of cell system have rarely been incorporated into the existing algorithms for detecting protein complexes because of the limitation of protein-protein interaction (PPI) data produced by high throughput techniques. The availability of time course gene expression profile enables us to uncover the dynamics of molecular networks and improve the detection of protein complexes...
2017: PloS One
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