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Tubulin

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https://www.readbyqxmd.com/read/28448000/high-resolution-imaging-and-analysis-of-individual-astral-microtubule-dynamics-in-budding-yeast
#1
Colby P Fees, Cassi Estrem, Jeffrey K Moore
Dynamic microtubules are fundamental to many cellular processes, and accurate measurements of microtubule dynamics can provide insight into how cells regulate these processes and how genetic mutations impact regulation. The quantification of microtubule dynamics in metazoan models has a number of associated challenges, including a high microtubule density and limitations on genetic manipulations. In contrast, the budding yeast model offers advantages that overcome these challenges. This protocol describes a method to measure the dynamics of single microtubules in living yeast cells...
April 20, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28444612/wbp2nl-pawp-mrna-and-protein-expression-in-sperm-cells-are-not-related-to-semen-parameters-fertilization-rate-or-reproductive-outcome
#2
T Freour, M Barragan, A Ferrer-Vaquer, A Rodríguez, Rita Vassena
PURPOSE: WBP2NL/PAWP, a protein found in the post-acrosomal region of mammalian spermatozoa, has been proposed as a sperm-borne oocyte-activating factor (SOAF) contributing to Ca(2+) release within the oocyte and subsequent fertilization and embryo development. However, its relevance as either a diagnostic or a prognostic marker of fertilization failure has been questioned in the recent literature. We analyzed WBP2NL/PAWP gene and protein expression level and localization in patients without previous intracytoplasmic sperm injection (ICSI) cycles in order to assess its association with both sperm characteristics and ability to fertilize...
March 21, 2017: Journal of Assisted Reproduction and Genetics
https://www.readbyqxmd.com/read/28441930/ribosomal-subunit-protein-typing-using-matrix-assisted-laser-desorption-ionization-time-of-flight-mass-spectrometry-maldi-tof-ms-for-the-identification-and-discrimination-of-aspergillus-species
#3
Sayaka Nakamura, Hiroaki Sato, Reiko Tanaka, Yoko Kusuya, Hiroki Takahashi, Takashi Yaguchi
BACKGROUND: Accurate identification of Aspergillus species is a very important subject. Mass spectral fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is generally employed for the rapid identification of fungal isolates. However, the results are based on simple mass spectral pattern-matching, with no peak assignment and no taxonomic input. We propose here a ribosomal subunit protein (RSP) typing technique using MALDI-TOF MS for the identification and discrimination of Aspergillus species...
April 26, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28441562/bacterial-division-ftsz-treadmills-to-build-a-beautiful-wall
#4
Kara M Schoenemann, William Margolin
The tubulin-like FtsZ protein polymerizes into a contractile ring structure required for cytokinesis in most bacteria. Two new studies report that FtsZ polymers move around the ring by treadmilling, which guides and regulates the inward growth of the septal wall.
April 24, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28440465/5-furan-2-yl-4-3-4-5-trimethoxyphenyl-3h-1-2-dithiol-3-one-oxime-6f-a-new-synthetic-compound-causes-human-%C3%AF-brosarcoma-ht-1080-cell-apoptosis-by-disrupting-tubulin-polymerisation-and-inducing-g2-m-arrest
#5
Daiying Zuo, Lili Pang, Jiwei Shen, Qi Guan, Zhaoshi Bai, Huijuan Zhang, Yao Li, Guodong Lu, Weige Zhang, Yingliang Wu
In the current study, we synthesized a series of new compounds targeting tubulin and tested their anti-proliferative activities. Among these new synthetic com-pounds, 5-(furan-2-yl)-4-(3,4,5-trimethoxyphenyl)-3H-1,2-dithiol-3-one oxime (6f) exhibited significant anti-proliferative activity against different human cancer cell lines including human gastric adenocarcinoma SGC-7901, human non-small cell lung cancer A549, and human fibrosarcoma HT-1080. As a result, 6f was selected to further test the sensitivity to different cancer cell lines including human cervical cancer cell line HeLa, human breast cancer cell line MCF-7, non-small cell lung cancer cell line A549, human liver carcinoma cell line HepG-2, human oral squamous cell carcinoma cell lines KB, SGC-7901 and HT-1080...
April 18, 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28438890/short-ftsz-filaments-can-drive-asymmetric-cell-envelope-constriction-at-the-onset-of-bacterial-cytokinesis
#6
Qing Yao, Andrew I Jewett, Yi-Wei Chang, Catherine M Oikonomou, Morgan Beeby, Cristina V Iancu, Ariane Briegel, Debnath Ghosal, Grant J Jensen
FtsZ, the bacterial homologue of eukaryotic tubulin, plays a central role in cell division in nearly all bacteria and many archaea. It forms filaments under the cytoplasmic membrane at the division site where, together with other proteins it recruits, it drives peptidoglycan synthesis and constricts the cell. Despite extensive study, the arrangement of FtsZ filaments and their role in division continue to be debated. Here, we apply electron cryotomography to image the native structure of intact dividing cells and show that constriction in a variety of Gram-negative bacterial cells, including Proteus mirabilis and Caulobacter crescentus, initiates asymmetrically, accompanied by asymmetric peptidoglycan incorporation and short FtsZ-like filament formation...
April 24, 2017: EMBO Journal
https://www.readbyqxmd.com/read/28437075/fluorescent-photoswitchable-diarylethenes-for-biolabeling-and-single-molecule-localization-microscopies-with-optical-superresolution
#7
Benoit Roubinet, Michael Weber, Heydar Shojaei, Mark Bates, Mariano L Bossi, Vladimir N Belov, Masahiro Irie, Stefan W Hell
A modular assembly of water soluble diarylethenes (DAEs), applicable as bio-markers for optical nanoscopy, is reported. Reversibly photoswitchable 1,2-bis(2-alkyl-6-phenyl-1-benzothiophene-1,1-dioxide-3-yl)perfluorocyclopentenes possessing a fluorescent "closed" form were decorated with one or two methoxy group(s) attached to the para-position(s) of phenyl ring(s) and two, four or eight carboxylic acid groups. Antibody conjugates of these DAEs feature low aggregation, efficient photoswitching in aqueous buffers, specific staining of cellular structures and photophysical properties (high emission efficiencies and low cycloreversion quantum yields) enabling their application in superresolution microscopy...
April 24, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28436240/biotherapeutic-potential-and-mechanisms-of-action-of-colchicine
#8
Kashyap Kumar Dubey, Punit Kumar, Nikolaos E Labrou, Pratyoosh Shukla
Cancer is a clinical situation caused by uncontrolled cell division and is responsible for a large number of deaths worldwide. Colchicine is a classical antimitotic, tubulin-binding agent (TBA) which is being explored for its antitumor activities, although its tubulin-binding ability leads to some toxicity toward normal cells proliferation. Colchicine derivatives are considered as potent antitumor compounds with less toxicity compared to colchicine. Derivatives with substituted functional groups at A-ring (methoxy), B-ring (acetamide) or C-ring (methoxy) have been synthesized via chemical and microbial routes and show modified bioactivities and altered tropolonic functionality...
April 23, 2017: Critical Reviews in Biotechnology
https://www.readbyqxmd.com/read/28436114/all-trans-retinoic-acid-stimulates-overexpression-of-tumor-protein-d52-tpd52-isoform-3-and-neuronal-differentiation-of-imr-32-cells
#9
Sudha Sravanti Kotapalli, Chandrashekhar Dasari, Divya Duscharla, Karthik Reddy Kami Reddy, Manjula Kasula, Ramesh Ummanni
Tumor protein D52 (TPD52), a proto-oncogene is overexpressed in a variety of epithelial carcinomas and plays an important role in cell proliferation, migration and cell death. In the present study we found that the treatment of IMR-32 neuroblastoma (NB) cells with retinoic acid (RA) stimulates an increase in expression of TPD52. TPD52 expression is detectable after 72 h, can be maintained till differentiation of NB cells suggesting that TPD52 is involved in differentiation. Here, we demonstrate that TPD52 is essential for RA to promote differentiation of NB cells...
April 24, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28435525/development-and-biological-evaluation-of-a-photoactivatable-small-molecule-microtubule-targeting-agent
#10
Alexander Döbber, Athena F Phoa, Ramzi H Abbassi, Brett W Stringer, Bryan W Day, Terrance G Johns, Mohammed Abadleh, Christian Peifer, Lenka Munoz
Photoremovable protecting groups added to bioactive molecules provide spatial and temporal control of the biological effects. We present synthesis and characterization of the first photoactivatable small-molecule tubulin inhibitor. By blocking the pharmacophoric OH group on compound 1 with photoremovable 4,5-dimethoxy-2-nitrobenzyl moiety we developed the photocaged prodrug 2 that had no effect in biological assays. Short UV light exposure of the derivative 2 or UV-irradiation of cells treated with 2 resulted in fast and potent inhibition of tubulin polymerization, attenuation of cell viability, and apoptotic cell death, implicating release of the parent active compound...
April 13, 2017: ACS Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28434791/neuronal-cells-derived-from-human-induced-pluripotent-stem-cells-as-a-functional-tool-of-melanocortin-system
#11
Nobuko Yamada-Goto, Yukari Ochi, Goro Katsuura, Yui Yamashita, Ken Ebihara, Michio Noguchi, Junji Fujikura, Daisuke Taura, Masakatsu Sone, Kiminori Hosoda, Paul E Gottschall, Kazuwa Nakao
BACKGROUND: The preparation of human neurons derived from human induced pluripotent stem (iPS) cells can serve as a potential tool for evaluating the physiological and pathophysiological properties of human neurons and for drug development. METHODS: In the present study, the functional activity in neuronal cells differentiated from human iPS cells was observed. RESULTS: The differentiated cells expressed mRNAs for classical neuronal markers (microtubule-associated protein 2, β-tubulin III, calbindin 1, synaptophysin and postsynaptic density protein 95) and for subunits of various excitatory and inhibitory transmitters (NR1, NR2A, NR2B, GABAA α1)...
April 7, 2017: Neuropeptides
https://www.readbyqxmd.com/read/28433265/high-pressure-microscopy-for-tracking-dynamic-properties-of-molecular-machines
#12
Masayoshi Nishiyama
High-pressure microscopy is one of the powerful techniques to visualize the effects of hydrostatic pressures on research targets. It could be used for monitoring the pressure-induced changes in the structure and function of molecular machines in vitro and in vivo. This review focuses on the dynamic properties of the assemblies and machines, analyzed by means of high-pressure microscopy measurement. We developed a high-pressure microscope that is optimized both for the best image formation and for the stability to hydrostatic pressure up to 150 MPa...
March 30, 2017: Biophysical Chemistry
https://www.readbyqxmd.com/read/28431353/discovery-of-potential-anticancer-multi-targeted-ligustrazine-based-cyclohexanone-and-oxime-analogs-overcoming-the-cancer-multidrug-resistance
#13
Gao-Feng Zha, Hua-Li Qin, Bahaa G M Youssif, Muhammad Wahab Amjad, Maria Abdul Ghafoor Raja, Ahmed H Abdelazeem, Syed Nasir Abbas Bukhari
The drug research and development nowadays is focusing on multi-target drugs. In the treatment of cancer, therapies using drugs inhibiting one numerous targets signify a novel viewpoint. In comparison with traditional therapy, multi-targeted drugs directly aim cell subpopulations which are involved in progression of tumor. The current study comprises the synthesis of 34 novel ligustrazine-containing α, β-unsaturated carbonyl-based compounds and oximes. The growth of 5 various cancer cell types was strongly inhibited by ligustrazine-containing oximes as revealed by biological evaluation...
April 14, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28430413/bioimaging-with-macromolecular-probes-incorporating-multiple-bodipy-fluorophores
#14
Ek Raj Thapaliya, Yang Zhang, Pravat Dhakal, Adrienne S Brown, James N Wilson, Kevin Collins, Françisco M Raymo
Seven macromolecular constructs incorporating multiple borondipyrromethene (BODIPY) fluorophores along a common poly(methacrylate) backbone with decyl and oligo(ethylene glycol) side chains were synthesized. The hydrophilic oligo(ethylene glycol) components impose solubility in aqueous environment on the overall assembly. The hydrophobic decyl chains insulate effectively the fluorophores from each other to prevent detrimental interchromophoric interactions and preserve their photophysical properties. As a result, the brightness of these multicomponent assemblies is approximately three times greater than that of a model BODIPY monomer...
April 21, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28429320/euglena-gracilis-genome-and-transcriptome-organelles-nuclear-genome-assembly-strategies-and-initial-features
#15
ThankGod Echezona Ebenezer, Mark Carrington, Michael Lebert, Steven Kelly, Mark C Field
Euglena gracilis is a major component of the aquatic ecosystem and together with closely related species, is ubiquitous worldwide. Euglenoids are an important group of protists, possessing a secondarily acquired plastid and are relatives to the Kinetoplastidae, which themselves have global impact as disease agents. To understand the biology of E. gracilis, as well as to provide further insight into the evolution and origins of the Kinetoplastidae, we embarked on sequencing the nuclear genome; the plastid and mitochondrial genomes are already in the public domain...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28428427/microtubules-acquire-resistance-from-mechanical-breakage-through-intralumenal-acetylation
#16
Zhenjie Xu, Laura Schaedel, Didier Portran, Andrea Aguilar, Jérémie Gaillard, M Peter Marinkovich, Manuel Théry, Maxence V Nachury
Eukaryotic cells rely on long-lived microtubules for intracellular transport and as compression-bearing elements. We considered that long-lived microtubules are acetylated inside their lumen and that microtubule acetylation may modify microtubule mechanics. Here, we found that tubulin acetylation is required for the mechanical stabilization of long-lived microtubules in cells. Depletion of the tubulin acetyltransferase TAT1 led to a significant increase in the frequency of microtubule breakage. Nocodazole-resistant microtubules lost upon removal of acetylation were largely restored by either pharmacological or physical removal of compressive forces...
April 21, 2017: Science
https://www.readbyqxmd.com/read/28428042/synthesis-and-biological-evaluation-of-selective-tubulin-inhibitors-as-anti-trypanosomal-agents
#17
Viharika Bobba, Vishal Nanavaty, Nethrie D Idippily, Anran Zhao, Bibo Li, Bin Su
African trypanosomiasis is still a threat to human health due to the severe side-effects of current drugs. We identified selective tubulin inhibitors that showed the promise to the treatment of this disease, which was based on the tubulin protein structural difference between mammalian and trypanosome cells. Further lead optimization was performed in the current study to improve the efficiency of the drug candidates. We used Trypanosoma brucei brucei cells as the parasite model, and human normal kidney cells and mouse macrophage cells as the host model to evaluate the compounds...
April 8, 2017: Bioorganic & Medicinal Chemistry
https://www.readbyqxmd.com/read/28427819/size-dependent-axial-instability-of-microtubules-surrounded-by-cytoplasm-of-a-living-cell-based-on-nonlocal-strain-gradient-elasticity-theory
#18
S Sahmani, M M Aghdam
Microtubules including tubulin heterodimers arranging in a parallel shape of cylindrical hollow plays an important role in the mechanical stiffness of a living cell. In the present study, the nonlocal strain gradient theory of elasticity including simultaneously the both nonlocality and strain gradient size dependency is put to use within the framework of a refined orthotropic shell theory with hyperbolic distribution of shear deformation to analyze the size-dependent buckling and postbuckling characteristics of microtubules embedded in cytoplasm under axial compressive load...
April 17, 2017: Journal of Theoretical Biology
https://www.readbyqxmd.com/read/28426223/over-destabilization-of-protein-protein-interaction-in-generalized-born-model-and-utility-of-energy-density-integration-cutoff
#19
Yukinobu Mizuhara, Dan Parkin, Koji Umezawa, Jun Ohnuki, Mitsunori Takano
The generalize Born (GB) model is frequently used in MD simulations of biomolecular systems in aqueous solution. The GB model is usually based on the so-called Coulomb field approximation (CFA) for the energy density integration. In this study, we report that the GB model with CFA overdestabilizes the long-range electrostatic attraction between oppositely-charged molecules (ionic bond forming two-helix system and kinesin-tubulin system) when the energy density integration cutoff, rmax, which is used to calculate the Born energy, is set to a large value...
April 20, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28425876/characterization-of-the-translation-elongation-factor-1-%C3%AE-gene-in-a-wide-range-of-pathogenic-aspergillus-species
#20
Sadegh Nouripour-Sisakht, Bahram Ahmadi, Koichi Makimura, Sybren de Hoog, Yoshiko Umeda, Mohamed Mahdi Alshahni, Hossein Mirhendi
PURPOSE: We aimed to evaluate the resolving power of the translation elongation factor (TEF)-1α gene for phylogenetic analysis of Aspergillus species. METHODOLOGY: Sequences of 526 bp representing the coding region of the TEF-1α gene were used for the assessment of levels of intra- and inter-specific nucleotide polymorphism in 33 species of Aspergillus, including 57 reference, clinical and environmental strains. RESULTS: Analysis of TEF-1α sequences indicated a mean similarity of 92...
April 21, 2017: Journal of Medical Microbiology
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