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rhodopsin

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https://www.readbyqxmd.com/read/29330515/hyphatracker-an-imagej-toolbox-for-time-resolved-analysis-of-spore-germination-in-filamentous-fungi
#1
Michael Brunk, Sebastian Sputh, Sören Doose, Sebastian van de Linde, Ulrich Terpitz
The dynamics of early fungal development and its interference with physiological signals and environmental factors is yet poorly understood. Especially computational analysis tools for the evaluation of the process of early spore germination and germ tube formation are still lacking. For the time-resolved analysis of conidia germination of the filamentous ascomycete Fusarium fujikuroi we developed a straightforward toolbox implemented in ImageJ. It allows for processing of microscopic acquisitions (movies) of conidial germination starting with drift correction and data reduction prior to germling analysis...
January 12, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29324661/protein-activity-of-the-fusarium-fujikuroi-rhodopsins-caro-and-opsa-and-their-relation-to-fungus-plant-interaction
#2
Alexander Adam, Stephan Deimel, Javier Pardo-Medina, Jorge García-Martínez, Tilen Konte, M Carmen Limón, Javier Avalos, Ulrich Terpitz
Fungi possess diverse photosensory proteins that allow them to perceive different light wavelengths and to adapt to changing light conditions in their environment. The biological and physiological roles of the green light-sensing rhodopsins in fungi are not yet resolved. The rice plant pathogen Fusarium fujikuroi exhibits two different rhodopsins, CarO and OpsA. CarO was previously characterized as a light-driven proton pump. We further analyzed the pumping behavior of CarO by patch-clamp experiments. Our data show that CarO pumping activity is strongly augmented in the presence of the plant hormone indole-3-acetic acid and in sodium acetate, in a dose-dependent manner under slightly acidic conditions...
January 11, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29320632/increasing-the-stability-of-recombinant-human-green-cone-pigment
#3
Timothy S Owen, David Salom, Wenyu Sun, Krzysztof Palczewski
Three types of cone cells exist in the human retina, each containing a different pigment responsible for the initial step of photo-transduction. These pigments are distinguished by their specific absorbance maxima, 425 nm (blue), 530 nm (green) and 560 nm (red). Each pigment contains a common chromophore, 11-cis-retinal covalently bound to an opsin protein via a Schiff base. The 11-cis-retinal protonated Schiff base has an absorbance maxima at 440 nm in methanol. Unfortunately, the chemistry allowing the same chromophore interacting with different opsin proteins to tune the absorbance of the resulting pigments to distinct λmax values is poorly understood...
January 10, 2018: Biochemistry
https://www.readbyqxmd.com/read/29315318/correlative-light-and-immuno-electron-microscopy-of-retinal-tissue-cryostat-sections
#4
Thomas Burgoyne, Amelia Lane, William E Laughlin, Michael E Cheetham, Clare E Futter
Correlative light-electron microscopy (CLEM) is a powerful technique allowing localisation of specific macromolecules within fluorescence microscopy (FM) images to be mapped onto corresponding high-resolution electron microscopy (EM) images. Existing methods are applicable to limited sample types and are technically challenging. Here we describe novel methods to perform CLEM and immuno-electron microscopy (iEM) on cryostat sections utilising the popular FM embedding solution, optimal cutting temperature (OCT) compound...
2018: PloS One
https://www.readbyqxmd.com/read/29311491/visual-evoked-potential-in-rabbits-eyes-with-subretinal-implantation-by-vitrectomy-of-okayama-university-type-retinal-prosthesis-oureptm
#5
Toshihiko Matsuo, Tetsuya Uchida, Koichiro Yamashita, Shigiko Takei, Daisuke Ido, Mamoru Tanaka, Masao Oguchi, Toshinori Furukawa
Okayama University-type retinal prosthesis (OURePTM) is a photoelectric dye-coupled polyethylene film which generates electric potential in response to light and stimulates nearby neurons. This study aims to test surgical feasibility for subretinal film implantation and to examine functional durability of films in subretinal space. Dye-coupled films were implanted subretinally by vitrectomy in the right eye of normal white rabbits: 8 rabbits for 1 month and 8 rabbits for 6 months. The implanted films were removed by vitrectomy in 4 of these 8 rabbits in 1-month or 6-month implantation group...
January 1, 2018: Journal of Veterinary Medical Science
https://www.readbyqxmd.com/read/29305604/molecular-defects-of-the-disease-causing-human-arrestin-1-c147f-mutant
#6
Sergey A Vishnivetskiy, Lori S Sullivan, Sara J Bowne, Stephen P Daiger, Eugenia V Gurevich, Vsevolod V Gurevich
Purpose: The purpose of this study was to identify the molecular defect in the disease-causing human arrestin-1 C147F mutant. Methods: The binding of wild-type (WT) human arrestin-1 and several mutants with substitutions in position 147 (including C147F, which causes dominant retinitis pigmentosa in humans) to phosphorylated and unphosphorylated light-activated rhodopsin was determined. Thermal stability of WT and mutant human arrestin-1, as well as unfolded protein response in 661W cells, were also evaluated...
January 1, 2018: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/29281027/cas9-sgrna-selective-targeting-of-the-p23h-rhodopsin-mutant-allele-for-treating-retinitis-pigmentosa-by-intravitreal-aav9-php-b-based-delivery
#7
Serena G Giannelli, Mirko Luoni, Valerio Castoldi, Luca Massimino, Tommaso Cabassi, Debora Angeloni, Giancarlo Demontis, Letizia Leocani, Massimiliano Andreazzoli, Vania Broccoli
P23H is the most common mutation in the RHODOPSIN (RHO) gene leading to a dominant form of Retinitis Pigmentosa (RP), a rod photoreceptor degeneration that invariably causes vision loss. Specific disruption of the disease P23H RHO mutant while preserving the wild-type functional allele would be an invaluable therapy for this disease. However, various technologies tested in the past failed to achieve effective changes and consequently therapeutic benefits. We validated a CRISPR/Cas9 strategy to specifically inactivate the P23H RHO mutant, while preserving the wild-type (WT) allele in vitro...
December 21, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/29274337/generation-of-three-dimensional-retinal-organoids-expressing-rhodopsin-and-s-and-m-cone-opsins-from-mouse-stem-cells
#8
Kaori Ueda, Akishi Onishi, Shin-Ichiro Ito, Makoto Nakamura, Masayo Takahashi
PURPOSE: Three-dimensional retinal organoids can be differentiated from embryonic stem cells/induced pluripotent stem cells (ES/iPS cells) under defined medium conditions. We modified the serum-free floating culture of embryoid body-like aggregates with quick reaggregation (SFEBq) culture procedure to obtain retinal organoids expressing more rod photoreceptors and S- and M-cone opsins. METHODS: Retinal organoids differentiated from mouse Nrl-eGFP iPS cells were cultured in various mediums during photoreceptor development...
December 20, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29242588/in-retinitis-pigmentosa-trkc-t1-dependent-vectorial-erk-activity-upregulates-glial-tnf-%C3%AE-causing-selective-neuronal-death
#9
Alba Galán, Sean Jmaeff, Pablo F Barcelona, Fouad Brahimi, Marinko V Sarunic, H Uri Saragovi
In some diseases the TrkC.T1 isoform is upregulated in glia, associated with glial TNF-α production and neuronal death. What remains unknown are the activating signals in glia, and how paracrine signals may be selective for a targeted neuron while sparing other proximate neurons. We studied these questions in the retina, where Müller glia contacts photoreceptors on one side and retinal ganglion cells on the other. In a mutant Rhodopsin mouse model of retinitis pigmentosa (RP) causing progressive photoreceptor death-but sparing retinal ganglion cells-TrkC...
December 14, 2017: Cell Death & Disease
https://www.readbyqxmd.com/read/29238742/class-c-g-protein-coupled-receptors-reviving-old-couples-with-new-partners
#10
Thor C Møller, David Moreno-Delgado, Jean-Philippe Pin, Julie Kniazeff
G protein-coupled receptors (GPCRs) are key players in cell communication and are encoded by the largest family in our genome. As such, GPCRs represent the main targets in drug development programs. Sequence analysis revealed several classes of GPCRs: the class A rhodopsin-like receptors represent the majority, the class B includes the secretin-like and adhesion GPCRs, the class F includes the frizzled receptors, and the class C includes receptors for the main neurotransmitters, glutamate and GABA, and those for sweet and umami taste and calcium receptors...
2017: Biophysics Reports
https://www.readbyqxmd.com/read/29232624/genomic-form-of-rhodopsin-dna-nanoparticles-rescued-autosomal-dominant-retinitis-pigmentosa-in-the-p23h-knock-in-mouse-model
#11
Rajendra Narayan Mitra, Min Zheng, Ellen R Weiss, Zongchao Han
Retinitis pigmentosa (RP) is a group of inherited retinal degenerative conditions and a leading cause of irreversible blindness. 25%-30% of RP cases are caused by inherited autosomal dominant (ad) mutations in the rhodopsin (Rho) protein of the retina, which impose a barrier for developing therapeutic treatments for this genetically heterogeneous disorder, as simple gene replacement is not sufficient to overcome dominant disease alleles. Previously, we have explored using the genomic short-form of Rho (sgRho) for gene augmentation therapy of RP in a Rho knockout mouse model...
December 5, 2017: Biomaterials
https://www.readbyqxmd.com/read/29228925/daily-activity-patterns-influence-retinal-morphology-signatures-of-selection-and-spectral-tuning-of-opsin-genes-in-colubrid-snakes
#12
E Hauzman, D M O Bonci, E Y Suárez-Villota, M Neitz, D F Ventura
BACKGROUND: Morphological divergences of snake retinal structure point to complex evolutionary processes and adaptations. The Colubridae family has a remarkable variety of retinal structure that can range from all-cone and all-rod to duplex (cone/rod) retinas. To explore whether nocturnal versus diurnal activity is responsible for constraints on molecular evolution and plays a role in visual opsin spectral tuning of colubrids, we carried out molecular evolution analyses of the visual opsin genes LWS, RH1, and SWS1 from 17 species and performed morphological analyses...
December 11, 2017: BMC Evolutionary Biology
https://www.readbyqxmd.com/read/29220051/improving-extraction-and-post-purification-concentration-of-membrane-proteins
#13
Hasin Feroz, HyeYoung Kwon, Jing Peng, Hyeonji Oh, Bryan Ferlez, Carol S Baker, John H Golbeck, Guillermo C Bazan, Andrew L Zydney, Manish Kumar
Membrane proteins (MPs), despite being critically important drug targets for the pharmaceutical industry, are difficult to study due to challenges in obtaining high yields of functional protein. Most current extraction efforts use specialized non-ionic detergents to solubilize and stabilize MPs, with MPs being concentrated by ultrafiltration (UF). However, many detergents are retained during the UF step, which can destabilize MPs and/or interfere with their characterization. Here, we studied the influence of detergent selection on the extraction and UF-based concentration of biomedically-relevant MPs, the light-driven sodium and chloride transporters, KR2 and halorhodopsin (pHR) which are also model proteins for more complex mammalian rhodopsins...
December 8, 2017: Analyst
https://www.readbyqxmd.com/read/29215887/unique-hydrogen-bonds-in-membrane-protein-monitored-by-whole-mid-ir-atr-spectroscopy-in-aqueous-solution
#14
Shota Ito, Masayo Iwaki, Shinya Sugita, Rei Abe-Yoshizumi, Tatsuya Iwata, Keiichi Inoue, Hideki Kandori
Protein function is coupled to its structural changes, for which stimulus-induced difference Fourier-transform infrared (FTIR) spectroscopy is a powerful method. By optimizing the attenuated tonal reflection (ATR)-FTIR analysis on the sodium pumping rhodopsin KR2 in aqueous solution, we first measured the accurate difference spectra upon sodium binding in the whole IR region (4000-1000 cm-1). The new spectral window allows the analysis of not only the finger print region (1800-1000 cm-1), but also the hydrogen-bonding donor region (4000-1800 cm-1), revealing an unusually strong hydrogen bond of Tyr located in the sodium binding site of KR2...
December 7, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/29212391/gene-therapy-successfully-delays-degeneration-in-a-mouse-model-of-pde6a-linked-retinitis-pigmentosa-rp-43
#15
Christian Schön, Vithiyanjali Sothilingam, Regine Mühlfriedel, Marina Garcia Garrido, Susanne C Beck, Naoyuki Tanimoto, Bernd Wissinger, François Paquet-Durand, Martin Biel, Stylianos Michalakis, Mathias W Seeliger, Rd-Cure Consortium
Retinitis pigmentosa type 43 (RP43) is a blinding disease caused by mutations in the gene for rod phosphodiesterase 6 alpha (PDE6A). The disease process begins with a dysfunction of rod photoreceptors, subsequently followed by a currently untreatable progressive degeneration of the entire outer retina. Aiming at a curative approach via PDE6A gene supplementation, we developed a novel adeno-associated viral (AAV) vector for expression of the human PDE6A cDNA under control of the human rhodopsin promotor (rAAV8...
December 7, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/29212382/gene-supplementation-rescues-rod-function-and-preserves-photoreceptor-and-retinal-morphology-in-dogs-leading-the-way-towards-treating-human-pde6a-retinitis-pigmentosa
#16
Laurence M Occelli, Christian Schön, Mathias W Seeliger, Martin Biel, Stylianos Michalakis, Simon Petersen-Jones, Rd-Cure Consortium
Mutations in the phosphodiesterase 6A gene (PDE6A) result in retinitis pigmentosa type 43 (RP43) and are responsible for about 4% of autosomal recessive RP. There is currently no treatment for this blinding condition. The aim of this project was to use a large animal model to test a gene supplementation viral vector designed to be translated for use in a clinical trial for the treatment of RP43. Seven Pde6a<sup>-/-</sup> puppies were given subretinal injections of an adeno-associated viral vector serotype 2/8 delivering human PDE6A cDNA under control of a short rhodopsin promoter (AAV8-PDE6A)...
December 6, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/29209002/the-orientation-and-stability-of-the-gpcr-arrestin-complex-in-a-lipid-bilayer
#17
Dali Wang, Hua Yu, Xiangdong Liu, Jianqiang Liu, Chen Song
G protein-coupled receptors (GPCRs) constitute a large family of membrane proteins that plays a key role in transmembrane signal transduction and draw wide attention since it was discovered. Arrestin is a small family of proteins which can bind to GPCRs, block G protein interactions and redirect signaling to G-protein-independent pathways. The detailed mechanism of how arrestin interacts with GPCR remains elusive. Here, we conducted molecular dynamics simulations with coarse-grained (CG) and all-atom (AA) models to study the complex structure formed by arrestin and rhodopsin, a prototypical GPCR, in a POPC bilayer...
December 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29203866/rpgrip1-is-required-for-rod-outer-segment-development-and-ciliary-protein-trafficking-in-zebrafish
#18
Rakesh K Raghupathy, Xun Zhang, Fei Liu, Reem H Alhasani, Lincoln Biswas, Saeed Akhtar, Luyuan Pan, Cecilia B Moens, Wenchang Li, Mugen Liu, Breandan N Kennedy, Xinhua Shu
Mutations in the RPGR-interacting protein 1 (RPGRIP1) gene cause recessive Leber congenital amaurosis (LCA), juvenile retinitis pigmentosa (RP) and cone-rod dystrophy. RPGRIP1 interacts with other retinal disease-causing proteins and has been proposed to have a role in ciliary protein transport; however, its function remains elusive. Here, we describe a new zebrafish model carrying a nonsense mutation in the rpgrip1 gene. Rpgrip1homozygous mutants do not form rod outer segments and display mislocalization of rhodopsin, suggesting a role for RPGRIP1 in rhodopsin-bearing vesicle trafficking...
December 4, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29188557/characterization-of-ligand-binding-to-gpcrs-through-computational-methods
#19
Silvana Vasile, Mauricio Esguerra, Willem Jespers, Ana Oliveira, Jessica Sallander, Johan Åqvist, Hugo Gutiérrez-de-Terán
The recent increase in available G protein-coupled receptor structures now contributes decisively to the structure-based ligand design. In this context, computational approaches in combination with medicinal chemistry and pharmacology are extremely helpful. Here, we provide an update on our structure-based computational protocols, used to answer key questions related to GPCR-ligand binding. All combined, these techniques can shed light on ligand binding modes, determine the molecular basis of conformational selection, for agonists and antagonists, as well as of subtype selectivity...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29188556/current-and-future-challenges-in-gpcr-drug-discovery
#20
Sid Topiol
GPCRs play a pervasive physiological role and, in turn, are the leading target class for pharmaceuticals. Beginning with the determination of the structure of rhodopsin, and dramatically accelerating since the reporting of the first ligand-mediated GPCR X-ray structures, our understanding of the structural and functional characteristics of these proteins has grown dramatically. Deploying this now rapidly emerging information for drug discovery has already been extensively demonstrated through a watershed of studies appearing in numerous scientific reports...
2018: Methods in Molecular Biology
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