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https://www.readbyqxmd.com/read/27913623/chac2-an-enzyme-for-slow-turnover-of-cytosolic-glutathione
#1
Amandeep Kaur, Ruchi Gautam, Ritika Srivastava, Avinash Chandel, Akhilesh Kumar, Subramanian Karthikeyan, Anand Kumar Bachhawat
Glutathione degradation plays an important role in glutathione and redox homeostasis and thus it is imperative to understand the enzymes and the mechanisms involved in glutathione degradation in detail. We describe here ChaC2, a member of the ChaC family of γ-glutamylcyclotransferases, as an enzyme that degrades glutathione in the cytosol of mammalian cells. ChaC2 is distinct from the previously described ChaC1, to which ChaC2 shows about 50% sequence identity. Human and mouse ChaC2 proteins, purified in vitro, show 10-20 fold lower catalytic efficiency than ChaC1, although they showed comparable Km values (Km of 3...
December 2, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27913418/a-novel-rhamnose-rich-hetero-exopolysaccharide-isolated-from-lactobacillus-paracasei-dg-activates-thp-1-human-monocytic-cells
#2
Silvia Balzaretti, Valentina Taverniti, Simone Guglielmetti, Walter Fiore, Mario Minuzzo, Hansel N Ngo, Judith B Ngere, Sohaib Sadiq, Paul N Humphreys, Andrew P Laws
: Lactobacillus paracasei DG is a bacterial strain with recognized probiotic properties and is used in commercial probiotic products. However, the mechanisms underlying its probiotic properties are mainly unknown. In this study, we tested the hypothesis that the capability of strain DG to interact with the host is, at least partly, associated with its ability to synthesize a surface-associated exopolysaccharide (EPS). Comparative genomics revealed the presence of putative EPS gene clusters in DG genome; accordingly, EPS was isolated from the surface of the bacterium...
December 2, 2016: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/27912755/establishment-of-high-reciprocal-connectivity-between-clonal-cortical-neurons-is-regulated-by-the-dnmt3b-dna-methyltransferase-and-clustered-protocadherins
#3
Etsuko Tarusawa, Makoto Sanbo, Atsushi Okayama, Toshio Miyashita, Takashi Kitsukawa, Teruyoshi Hirayama, Takahiro Hirabayashi, Sonoko Hasegawa, Ryosuke Kaneko, Shunsuke Toyoda, Toshihiro Kobayashi, Megumi Kato-Itoh, Hiromitsu Nakauchi, Masumi Hirabayashi, Takeshi Yagi, Yumiko Yoshimura
BACKGROUND: The specificity of synaptic connections is fundamental for proper neural circuit function. Specific neuronal connections that underlie information processing in the sensory cortex are initially established without sensory experiences to a considerable extent, and then the connections are individually refined through sensory experiences. Excitatory neurons arising from the same single progenitor cell are preferentially connected in the postnatal cortex, suggesting that cell lineage contributes to the initial wiring of neurons...
December 2, 2016: BMC Biology
https://www.readbyqxmd.com/read/27912176/novel-designed-vmct1-analogs-with-increased-antimicrobial-activity
#4
Cibele Nicolaski Pedron, Marcelo Der Torossian Torres, Julia Aparecida da Silva Lima, Pedro Ismael Silva, Fernanda Dias Silva, Vani Xavier Oliveira
Antimicrobial peptides are biologically active molecules produced by a wide range of organisms as an essential component of the innate immune response. They have recently attracted great interest, since they have antimicrobial activity against a broad spectrum of microorganisms. VmCT1 is a cationic peptide from the venom of Vaejovis mexicanus smithi scorpions, which presents antibacterial activity and tends to helical structures. Its analogs were synthesized, characterized and the conformational studies were performed by circular dichroism...
November 24, 2016: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/27911841/high-performance-multiplexed-fluorescence-in-situ-hybridization-in-culture-and-tissue-with-matrix-imprinting-and-clearing
#5
Jeffrey R Moffitt, Junjie Hao, Dhananjay Bambah-Mukku, Tian Lu, Catherine Dulac, Xiaowei Zhuang
Highly multiplexed single-molecule FISH has emerged as a promising approach to spatially resolved single-cell transcriptomics because of its ability to directly image and profile numerous RNA species in their native cellular context. However, background-from off-target binding of FISH probes and cellular autofluorescence-can become limiting in a number of important applications, such as increasing the degree of multiplexing, imaging shorter RNAs, and imaging tissue samples. Here, we developed a sample clearing approach for FISH measurements...
November 22, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27911811/the-structure-of-the-catechin-binding-site-of-human-sulfotransferase-1a1
#6
Ian Cook, Ting Wang, Mark Girvin, Thomas S Leyh
We are just beginning to understand the allosteric regulation of the human cytosolic sulfotransferase (SULTs) family-13 disease-relevant enzymes that regulate the activities of hundreds, if not thousands, of signaling small molecules. SULT1A1, the predominant isoform in adult liver, harbors two noninteracting allosteric sites, each of which binds a different molecular family: the catechins (naturally occurring flavonols) and nonsteroidal antiinflammatory drugs (NSAIDs). Here, we present the structure of an SULT allosteric binding site-the catechin-binding site of SULT1A1 bound to epigallocatechin gallate (EGCG)...
November 23, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27911791/a-functional-role-for-intrinsic-disorder-in-the-tau-tubulin-complex
#7
Ana M Melo, Juliana Coraor, Garrett Alpha-Cobb, Shana Elbaum-Garfinkle, Abhinav Nath, Elizabeth Rhoades
Tau is an intrinsically disordered protein with an important role in maintaining the dynamic instability of neuronal microtubules. Despite intensive study, a detailed understanding of the functional mechanism of tau is lacking. Here, we address this deficiency by using intramolecular single-molecule Förster Resonance Energy Transfer (smFRET) to characterize the conformational ensemble of tau bound to soluble tubulin heterodimers. Tau adopts an open conformation on binding tubulin, in which the long-range contacts between both termini and the microtubule binding region that characterize its compact solution structure are diminished...
November 23, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27911771/stability-folding-dynamics-and-long-range-conformational-transition-of-the-synaptic-t-snare-complex
#8
Xinming Zhang, Aleksander A Rebane, Lu Ma, Feng Li, Junyi Jiao, Hong Qu, Frederic Pincet, James E Rothman, Yongli Zhang
Synaptic soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) couple their stepwise folding to fusion of synaptic vesicles with plasma membranes. In this process, three SNAREs assemble into a stable four-helix bundle. Arguably, the first and rate-limiting step of SNARE assembly is the formation of an activated binary target (t)-SNARE complex on the target plasma membrane, which then zippers with the vesicle (v)-SNARE on the vesicle to drive membrane fusion. However, the t-SNARE complex readily misfolds, and its structure, stability, and dynamics are elusive...
November 28, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27911494/determining-functional-aptamer-protein-interaction-by-biolayer-interferometry
#9
Xinhui Lou, Martin Egli, Xianbin Yang
Short single-stranded nucleic acids called aptamers are widely being explored as recognition molecules of high affinity and specificity for binding a wide range of target molecules, particularly protein targets. In biolayer interferometry (BLI), a simple Dip-and-Read approach in which the aptamer-coated biosensors are dipped into microplate wells is used to study the interactions between an aptamer and its target protein. Here we describe the protocol for the analysis of the interaction between a well-characterized anti-thrombin RNA aptamer with thrombin (Basic Protocol)...
December 1, 2016: Current Protocols in Nucleic Acid Chemistry
https://www.readbyqxmd.com/read/27910974/fine-tuning-terminal-solvent-ligands-to-rationally-enhance-the-energy-barrier-in-dinuclear-dysprosium-single-molecule-magnets
#10
Kun Zhang, Chen Yuan, Fu-Sheng Guo, Yi-Quan Zhang, Yao-Yu Wang
In search of simple approaches to rationally enhance the energy barriers in polynuclear dysprosium single-molecule magnets, a new system containing two structurally closely related dinuclear dysprosium complexes, namely [Dy2(L)2(DBM)2(DMF)2] (1) and [Dy2(L)2(DBM)2(DMA)2]·2DMA (2) (HDBM = dibenzoylmethane, H2L = 2-hydroxy-N'-(2-hydroxy-3-methoxybenzylidene)benzohydrazide), is introduced and the structure-dependent magnetic properties are investigated. The two complexes display only slight variations in the coordination geometries of the Dy(iii) ion but display remarkably different magnetic behaviors...
December 2, 2016: Dalton Transactions: An International Journal of Inorganic Chemistry
https://www.readbyqxmd.com/read/27910874/comprehensive-evaluation-of-molecular-enhancers-of-the-isothermal-exponential-amplification-reaction
#11
Ellie Mok, Eugene Wee, Yuling Wang, Matt Trau
The exponential amplification reaction (EXPAR) is an emerging isothermal nucleic acid amplification method with high potential for molecular diagnostics due to its isothermal nature and high amplification efficiency. However, the use of EXPAR is limited by the high levels of non-specific amplification. Hence, methods that can improve the specificity of EXPAR are desired to facilitate its widespread adoption in practice. Herein, we proposed a strategy to improve EXPAR performance by using molecular enhancers...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27910521/a-low-temperature-external-cavity-diode-laser-for-broad-wavelength-tuning
#12
William G Tobias, Jason S Rosenberg, Nicholas R Hutzler, Kang-Kuen Ni
We report on the design and characterization of a low-temperature external cavity diode laser (ECDL) system for broad wavelength tuning. The performance achieved with multiple diode models addresses the scarcity of commercial red laser diodes below 633 nm, which is a wavelength range relevant to the spectroscopy of many molecules and ions. Using a combination of multiple-stage thermoelectric cooling and water cooling, the operating temperature of a laser diode is lowered to -64 °C, more than 85 °C below the ambient temperature...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27909986/toxicity-and-pharmacokinetic-profile-for-single-dose-injection-of-aby-029-a-fluorescent-anti-egfr-synthetic-affibody-molecule-for-human-use
#13
Kimberley S Samkoe, Jason R Gunn, Kayla Marra, Sally M Hull, Karen L Moodie, Joachim Feldwisch, Theresa V Strong, Daniel R Draney, P Jack Hoopes, David W Roberts, Keith Paulsen, Brian W Pogue
PURPOSE: ABY-029, a synthetic Affibody peptide, Z03115-Cys, labeled with a near-infrared fluorophore, IRDye® 800CW, targeting epidermal growth factor receptor (EGFR) has been produced under good manufacturing practices for a US Food and Drug Administration-approved first-in-use human study during surgical resection of glioma, as well as other tumors. Here, the pharmacology, phototoxicity, receptor activity, and biodistribution studies of ABY-029 were completed in rats, prior to the intended human use...
December 1, 2016: Molecular Imaging and Biology: MIB: the Official Publication of the Academy of Molecular Imaging
https://www.readbyqxmd.com/read/27909922/production-of-biologically-active-recombinant-goose-fsh-in-a-single-chain-form-with-a-ctp-linker-sequence
#14
Hui Li, Huanxi Zhu, Qinming Qin, Mingming Lei, Zhendan Shi
FSH is a glycoprotein hormone secreted by the pituitary gland that is essential for gonadal development and reproductive function. In avian reproduction study, especially in avian reproduction hormone study, it is hindered by the lack of biologically active FSH. In order to overcome this shortcoming, we prepared recombinant goose FSH as a single chain molecule and tested its biological activities in the present study. Coding sequences for mature peptides of goose FSH α and β subunits were amplified from goose pituitary cDNA...
December 1, 2016: Molecular Biology Reports
https://www.readbyqxmd.com/read/27908928/development-and-validation-of-a-normal-phase-chiral-hplc-method-for-analysis-of-afoxolaner-using-a-chiralpak%C3%A2-ad-3-column
#15
Jinyou Zhuang, Satish Kumar, Abu Rustum
Afoxolaner is a new antiparasitic molecule from the isoxazoline family that acts on the insect acarine gamma-aminobutyric acid and glutamate receptors. Isoxazoline family of compounds has been employed as active pharmaceutical ingredient in drug products prescribed for control of fleas and ticks in dogs. Afoxolaner with a chiral center at isoxazoline ring exists as a racemic mixture. A normal phase chiral high performance liquid chromatography analytical method has been developed to verify that afoxolaner is a racemic mixture as demonstrated by specific rotation, as well as to determine enantiomeric purity of single enantiomer samples...
November 2016: Journal of Chromatographic Science
https://www.readbyqxmd.com/read/27908126/communication-multipole-approximations-of-distant-pair-energies-in-local-correlation-methods-with-pair-natural-orbitals
#16
Hans-Joachim Werner
The accuracy of multipole approximations for distant pair energies in local second-order Møller-Plesset perturbation theory (LMP2) as introduced by Hetzer et al. [Chem. Phys. Lett. 290, 143 (1998)] is investigated for three chemical reactions involving molecules with up to 92 atoms. Various iterative and non-iterative approaches are compared, using different energy thresholds for distant pair selection. It is demonstrated that the simple non-iterative dipole-dipole approximation, which has been used in several recent pair natural orbitals (PNO)-LMP2 and PNO-LCCSD (local coupled-cluster with singles and doubles) methods, may underestimate the distant pair energies by up to 50% and can lead to significant errors in relative energies, unless very tight thresholds are used...
November 28, 2016: Journal of Chemical Physics
https://www.readbyqxmd.com/read/27906951/changes-in-neuroglial-activity-in-multiple-spinal-segments-after-caudal-epidural-pulsed-radiofrequency-in-a-rat-model-of-lumbar-disc-herniation
#17
Hee Kyung Cho, Jae Hoon Kang, So-Yeon Kim, Mi-Jung Choi, Se Jin Hwang, Yun-Woo Cho, Sang-Ho Ahn
BACKGROUND: Herniated lumbar discs can induce sciatica by mechanical compression and/or chemical irritation. It was recently reported that neuroglial cellular activity after pulsed radiofrequency (PRF) application to a single dorsal root ganglion (DRG) attenuated neuroglial activity at the corresponding spinal dorsal horn. Recently, caudal epidural PRF has been used to manage neuropathic pain, but evidence of molecular changes after the administration of caudal epidural PRF to attenuate neuropathic pain is lacking, and it has not been determined whether caudal epidural PRF affects neuroglial activity at different spinal levels...
November 2016: Pain Physician
https://www.readbyqxmd.com/read/27906373/efficient-homogeneous-illumination-and-optical-sectioning-for-quantitative-single-molecule-localization-microscopy
#18
Joran Deschamps, Andreas Rowald, Jonas Ries
Single-molecule localization microscopy (SMLM) relies on the switching of fluorescent molecules between a fluorescent and a dark state to achieve super resolution. This process is inherently dependent on the intensity distribution of the laser light used for both activation from the dark state and excitation of the bright state. Typically, laser light is coupled directly or via a single-mode fiber into the microscope, which leads to a Gaussian intensity profile in total internal reflection (TIR) or epi illumination...
November 28, 2016: Optics Express
https://www.readbyqxmd.com/read/27906230/stability-of-a-frequency-comb-based-transfer-lock-using-a-passive-fabry-perot-resonator
#19
Sambit B Pal, Mark M Lam, K Dieckmann
We report on a transfer-lock laser frequency stabilization that utilizes a frequency comb (FC) and a radio frequency counter referenced to a GPS frequency standard to compensate for the frequency drifts of two lasers, which are locked to a single passive Fabry-Perot resonator (FPR). The method requires only one optical phase lock with the FC and allows transfer locking of lasers at wavelengths beyond the usable range of the FC. To attain a large frequency tuning range for the lasers, we implement optical serrodyning...
December 1, 2016: Optics Letters
https://www.readbyqxmd.com/read/27906033/the-role-of-the-backbone-torsion-in-protein-folding
#20
EDITORIAL
Irina Sorokina, Arcady Mushegian
BACKGROUND: The set of forces and sequence of events that govern the transition from an unfolded polypeptide chain to a functional protein with correct spatial structure remain incompletely known, despite the importance of the problem and decades of theory development, computer simulations, and laboratory experiments. Information about the correctly folded state of most proteins is likely to be present in their sequences, and yet many proteins fail to attain native structure after overexpression in a non-native environment or upon experimental denaturation and refolding...
December 1, 2016: Biology Direct
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