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https://www.readbyqxmd.com/read/27924560/assaying-activation-and-subcellular-localization-of-erk-in-cells-and-tissues
#1
Carme Caelles, Carles Bayod, Melisa Morcillo
The extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) are the focus of many studies due to their involvement in numerous physiological and pathological processes, such as cell proliferation and differentiation, and oncogenic transformation, respectively. ERK1/2 belong to the mitogen-activated protein kinase (MAPKs) family, which are serine/threonine kinases that participate in signal transduction and are activated by dual phosphorylation in the Thr-X-Tyr motif located in their activation loop...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924260/molecular-mechanisms-of-substrate-specificities-of-uridinecytidine-kinase
#2
Wataru Tanaka, Mitsuo Shoji, Fumiaki Tomoike, Yuzuru Ujiie, Kyohei Hanaoka, Ryuhei Harada, Megumi Kayanuma, Katsumasa Kamiya, Toyokazu Ishida, Ryoji Masui, Seiki Kuramitsu, Yasuteru Shigeta
A uridine-cytidine kinase (UCK) catalyzes the phosphorylation of uridine (Urd) and cytidine (Cyd) and plays a significant role in the pyrimidine-nucleotide salvage pathway. Unlike ordinary ones, UCK from Thermus thermophilus HB8 (ttCK) loses catalytic activity on Urd due to lack of a substrate binding ability and possesses an unusual amino acid, i.e. tyrosine 93 (Tyr93) at the binding site, whereas histidine (His) is located in the other UCKs. Mutagenesis experiments revealed that a replacement of Tyr93 by His or glutamine (Gln) recovered catalytic activity on Urd...
2016: Biophysics and Physicobiology
https://www.readbyqxmd.com/read/27923741/nlrp3-inflammasome-activation-contributes-to-long-term-behavioral-alterations-in-mice-injected-with-lipopolysaccharide
#3
Wei Zhu, Feng-Sheng Cao, Jun Feng, Hua-Weng Chen, Jie-Ru Wan, Qing Lu, Jian Wang
Lipopolysaccharide (LPS) might affect the central nervous system by causing neuroinflammation, which subsequently leads to brain damage and dysfunction. In this study, we evaluated the role of Nod-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome activation in long-term behavioral alterations of 8-week-old male C57BL/6 mice injected intraperitoneally with LPS (5 mg/kg). At different time points after injection, we assessed locomotor function with a 24-point neurologic deficit scoring system and the rotarod test; assessed recognition memory with the novel object recognition test; and assessed emotional abnormality (anhedonia and behavioral despair) with the tail suspension test, forced swim test, and sucrose preference test...
December 3, 2016: Neuroscience
https://www.readbyqxmd.com/read/27919800/increasing-the-fidelity-of-noncanonical-amino-acid-incorporation-in-cell-free-protein-synthesis
#4
Qinglei Gan, Chenguang Fan
BACKGROUND: Cell-free protein synthesis provides a robust platform for co-translational incorporation of noncanonical amino acid (ncAA) into proteins to facilitate biological studies and biotechnological applications. Recently, eliminating the activity of release factor 1 has been shown to increase ncAA incorporation in response to amber codons. However, this approach could promote mis-incorporation of canonical amino acids by near cognate suppression. METHODS: We performed a facile protocol to remove near cognate tRNA isoacceptors of the amber codon from total tRNAs, and used the phosphoserine (Sep) incorporation system as validation...
December 2, 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27919619/microsatellite-polymorphism-located-immediately-upstream-of-the-phosphatidylinositol-glycan-class-k-gene-pigk-affects-its-expression-which-correlates-with-tyrosinase-activity-in-human-melanocytes
#5
Ken Okamura, Masahiro Hayashi, Yuko Abe, Yuta Araki, Yutaka Hozumi, Tamio Suzuki
BACKGROUND: Glycosylphosphatidylinositol (GPI) acts as a membrane anchor and a post-translational modifier for more than 150 proteins (called GPI-anchored proteins: GPI-APs). However, little study has been done to explore the role of GPI-APs in melanocytes. METHODS: The relationship between the mRNA expression of the genes which play essential roles in GPI anchoring system [phosphatidylinositol glycan, class A, and class K gene (PIGA, PIGK)] and melanogenesis-related genes (MITF, TYRP1, TYRP2, and TYR) as well as DOPA oxidase activities were evaluated in 13 different normal human epidermal melanocytes (NHEMs)...
November 27, 2016: Journal of Dermatological Science
https://www.readbyqxmd.com/read/27918933/tyrosine-fluorescence-probing-of-conformational-changes-in-tryptophan-lacking-domain-of-albumins
#6
N G Zhdanova, E G Maksimov, A M Arutyunyan, V V Fadeev, E A Shirshin
We addressed the possibility of using tyrosine (Tyr) fluorescence for monitoring conformational changes of proteins which are undetectable via tryptophan (Trp) fluorescence. The model objects, human (HSA) and bovine (BSA) serum albumins, contain one and two Trp residues, respectively, while Tyr is more uniformly distributed over their structure. The results of the investigation of albumins interaction with ethanol using intrinsic Trp and Tyr steady-state and time-resolved picosecond fluorescence indicated the presence of an intermediate at 10% (v/v) of ethanol in solution, that was supported by the results of extrinsic fluorescence measurements with the Nile Red dye...
November 25, 2016: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/27912144/elucidation-of-%C3%AE-s-dynamics-of-domain-iii-of-human-serum-albumin-during-the-chemical-and-thermal-unfolding-a-fluorescence-correlation-spectroscopic-investigation
#7
Bhaswati Sengupta, Nilimesh Das, Pratik Sen
The local structural dynamics and denaturation profile of domain-III of HSA against guanidine hydrochloride (GnHCl) and temperature has been studied using a coumarin based solvatochromic fluorescent probe p-nitrophenyl coumarin ester (NPCE), covalently tagged to Tyr-411 residue. By the steady state, time-resolved and single molecular level fluorescence studies it has been established that the domain-III of HSA is very sensitive to GnHCl but somewhat resistant to temperature and the domain specific unfolding proceeds in an altered way as compared to the overall unfolding of HSA...
November 18, 2016: Biophysical Chemistry
https://www.readbyqxmd.com/read/27911256/a-residue-located-at-the-junction-of-the-head-and-stalk-regions-of-measles-virus-fusion-protein-regulates-membrane-fusion-by-controlling-conformational-stability
#8
Yuto Satoh, Saeka Yonemori, Mitsuhiro Hirose, Hiroko Shogaki, Hiroshi Wakimoto, Yoshinori Kitagawa, Bin Gotoh, Tsuyoshi Shirai, Ken-Ichi Takahashi, Masae Itoh
The fusion (F) protein of measles virus (MV) performs refolding from the thermodynamically metastable prefusion form to the highly stable postfusion form via an activated unstable intermediate stage, to induce membrane fusion. Some amino acids involved in the fusion regulation cluster in the heptad repeat B (HR-B) domain of the stalk region, among which substitution of residue 465 by various amino acids revealed that fusion activity correlates well with its side chain length from the Cα (p<0.01) and van der Waals volume (p<0...
December 1, 2016: Journal of General Virology
https://www.readbyqxmd.com/read/27904871/in-vivo-measurement-and-characterization-of-a-novel-formulation-of-177-lu-dota-octreotate
#9
Dale L Bailey, Thomas M Hennessy, Kathy P Willowson, E Courtney Henry, David Lh Chan, Alireza Aslani, Paul J Roach
OBJECTIVES: Lutetium-177 can be made with high specific activity and with no other isotopes of lutetium present, referred to as "No Carrier Added" (NCA) (177)Lu. We have radiolabelled DOTA-conjugated peptide DOTA-(Tyr(3))-octreotate with NCA (177)Lu ("NCA-LuTATE") and used it in nearly 40 therapeutic administrations for subjects with neuroendocrine tumours or meningiomas. In this paper, we report on our initial studies on aspects of the biodistribution and dosimetry of NCA-LuTATE from gamma camera 2D whole body (WB) and quantitative 3D SPECT (qSPECT) (177)Lu imaging...
2016: Asia Oceania Journal of Nuclear Medicine & Biology
https://www.readbyqxmd.com/read/27904870/preclinical-study-of-68-ga-dotatoc-biodistribution-assessment-in-syrian-rats-and-evaluation-of-absorbed-dose-in-human-organs
#10
Mojdeh Naderi, Samaneh Zolghadri, Hassan Yousefnia, Ali Ramazani, Amir Reza Jalilian
OBJECTIVES: Gallium-68 DOTA-DPhe(1)-Tyr(3)-Octreotide ((68)Ga-DOTATOC) has been applied by several European centers for the treatment of a variety of human malignancies. Nevertheless, definitive dosimetric data are yet unavailable. According to the Society of Nuclear Medicine and Molecular Imaging, researchers are investigating the safety and efficacy of this radiotracer to meet Food and Drug Administration requirements. The aim of this study was to introduce the optimized procedure for (68)Ga-DOTATOC preparation, using a novel germanium-68 ((68)Ge)/(68)Ga generator in Iran and evaluate the absorbed doses in numerous organs with high accuracy...
2016: Asia Oceania Journal of Nuclear Medicine & Biology
https://www.readbyqxmd.com/read/27900439/non-ribosomal-peptide-synthases-from-pseudomonas-aeruginosa-play-a-role-in-cyclodipeptide-biosynthesis-quorum-sensing-regulation-and-root-development-in-a-plant-host
#11
Omar González, Randy Ortíz-Castro, César Díaz-Pérez, Alma L Díaz-Pérez, Viridiana Magaña-Dueñas, José López-Bucio, Jesús Campos-García
Diverse molecules mediate cross-kingdom communication between bacteria and their eukaryotic partners and determine pathogenic or symbiotic relationships. N-acyl-L-homoserine lactone-dependent quorum-sensing signaling represses the biosynthesis of bacterial cyclodipeptides (CDPs) that act as auxin signal mimics in the host plant Arabidopsis thaliana. In this work, we performed bioinformatics, biochemical, and plant growth analyses to identify non-ribosomal peptide synthase (NRPS) proteins of Pseudomonas aeruginosa, which are involved in CDP synthesis...
November 30, 2016: Microbial Ecology
https://www.readbyqxmd.com/read/27898860/effects-of-collection-time-on-flow-of-chromium-and-dry-matter-and-on-basal-ileal-endogenous-losses-of-amino-acids-in-growing-pigs
#12
B G Kim, Y Liu, H H Stein
The objectives of this experiment were to examine the diurnal patterns of chromium and DM flow at the distal ileum of pigs and to determine the effect of collection time on basal ileal endogenous losses (BEL) of CP and AA. Eight barrows with an initial BW of 34.6kg (SD = 2.1) were individually fitted with a T-cannula in the distal ileum and randomly allotted to a replicated 4× 4 Latin square design with 4 diets and 4 periods in each square. Three diets contained either corn, soybean meal, or distillers dried grains with solubles as the sole source of CP and AA...
October 2016: Journal of Animal Science
https://www.readbyqxmd.com/read/27892516/in-vivo-coherent-raman-imaging-of-the-melanomagenesis-associated-pigment-pheomelanin
#13
Hequn Wang, Sam Osseiran, Vivien Igras, Alexander J Nichols, Elisabeth M Roider, Joachim Pruessner, Hensin Tsao, David E Fisher, Conor L Evans
Melanoma is the most deadly form of skin cancer with a yearly global incidence over 232,000 patients. Individuals with fair skin and red hair exhibit the highest risk for developing melanoma, with evidence suggesting the red/blond pigment known as pheomelanin may elevate melanoma risk through both UV radiation-dependent and -independent mechanisms. Although the ability to identify, characterize, and monitor pheomelanin within skin is vital for improving our understanding of the underlying biology of these lesions, no tools exist for real-time, in vivo detection of the pigment...
November 28, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27888781/stabilization-of-cytochrome-b5-by-a-conserved-tyrosine-in-the-secondary-sphere-of-heme-active-site-a-spectroscopic-and-computational-study
#14
Shan Hu, Bo He, Xiao-Juan Wang, Shu-Qin Gao, Ge-Bo Wen, Ying-Wu Lin
Heme proteins perform a large array of biological functions, with the heme group bound non-covalently or covalently. To probe the stabilization role of conserved tyrosine residue in the secondary sphere of heme site in heme proteins, we herein used cytochrome b5 (Cyt b5) as a model protein, and mutated Tyr30 to Phe or His by removal of Tyr30 associated H-bond network and hydrophobic interaction. We performed thermal-induced unfolding studies for the two mutants, Y30F Cyt b5 and Y30H Cyt b5, as monitored by both UV-Vis and CD spectroscopy, as well as heme transfer studies from these proteins to apo-myoglobin, with wild-type Cyt b5 under the same conditions for comparison...
November 20, 2016: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/27886758/sensing-tyrosine-enantiomers-by-using-chiral-cdse-cds-quantum-dots-capped-with-n-acetyl-l-cysteine
#15
Fang Gao, Shiyao Ma, Xincai Xiao, Yan Hu, Dan Zhao, Zhike He
Despite of the importance of enantiomers, the fluorescence sensing of enantiomers and the interpretation of the "preferential interaction" still remain insufficiently explored. In this study, we report the recognition of tyrosine (Tyr) enantiomers by chiral N-acetyl-L-cysteine (L-NAC) capped CdSe/CdS quantum dots (QDs) under alkaline experimental condition. L-Tyr could greatly quench the fluorescence of CdSe/CdS QDs, while D-Tyr displayed no effect on the fluorescence. The one-step synthesized chiral L-NAC-CdSe/CdS QDs demonstrated high selectivity for Tyr enantiomers...
January 15, 2017: Talanta
https://www.readbyqxmd.com/read/27886256/structural-insights-into-the-interaction-of-the-ribosomal-p-stalk-protein-p2-with-a-type-ii-ribosome-inactivating-protein-ricin
#16
Xiaojiao Fan, Yuwei Zhu, Chongyuan Wang, Liwen Niu, Maikun Teng, Xu Li
Ricin is a type II ribosome-inactivating protein (RIP) that depurinates A(4324) at the sarcin-ricin loop of 28 S ribosomal RNA (rRNA), thus inactivating the ribosome by preventing elongation factors from binding to the GTPase activation centre. Recent studies have disclosed that the conserved C-terminal domain (CTD) of eukaryotic ribosomal P stalk proteins is involved in the process that RIPs target ribosome. However, the details of the molecular interaction between ricin and P stalk proteins remain unknown...
November 25, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27884696/design-synthesis-and-antifungal-activity-of-novel-fenfuram-diarylamine-hybrids
#17
Hongyu Wang, Xuheng Gao, Xiaoxiao Zhang, Hong Jin, Ke Tao, Taiping Hou
Ten novel fenfuram-diarylamine hybrids were designed and synthesized. And their antifungal activities against four phytopathogenic fungi have been evaluated in vitro and most of the compounds demonstrated a significant antifungal activities against Rhizoctonia solani and Sclerotinia sclerotiorum. Compound 5e exhibited the most potent antifungal activity against R. solani with an EC50 value of 0.037mg/L, far superior to the commercially available fungicide boscalid (EC50=1.71mg/L) and lead fungicide fenfuram (EC50=6...
November 12, 2016: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/27877847/adsorption-of-amino-acids-by-fullerenes-and-fullerene-nanowhiskers
#18
Hideo Hashizume, Chika Hirata, Kazuko Fujii, Kun'ichi Miyazawa
We have investigated the adsorption of some amino acids and an oligopeptide by fullerene (C60) and fullerene nanowhiskers (FNWs). C60 and FNWs hardly adsorbed amino acids. Most of the amino acids used have a hydrophobic side chain. Ala and Val, with an alkyl chain, were not adsorbed by the C60 or FNWs. Trp, Phe and Pro, with a cyclic structure, were not adsorbed by them either. The aromatic group of C60 did not interact with the side chain. The carboxyl or amino group, with the frame structure of an amino acid, has a positive or negative charge in solution...
December 2015: Science and Technology of Advanced Materials
https://www.readbyqxmd.com/read/27875310/extending-halogen-based-medicinal-chemistry-to-proteins-iodo-insulin-as-a-case-study
#19
Krystel El Hage, Vijay Pandyarajan, Nelson B Phillips, Brian J Smith, John G Menting, Jonathan Whittaker, Michael C Lawrence, Markus Meuwly, Michael A Weiss
Insulin, a protein critical for metabolic homeostasis, provides a classical model for protein design with application to human health. Recent efforts to improve its pharmaceutical formulation demonstrated that iodination of a conserved tyrosine (Tyr(B26)) enhances key properties of a rapid-acting clinical analog. Moreover, the broad utility of halogens in medicinal chemistry has motivated use of hybrid quantum- and molecular-mechanical methods to study proteins. Here, we (i) undertook quantitative atomic-level simulations of 3-I-Tyr(B26)-insulin to predict its structural features and (ii) tested these predictions by X-ray crystallography...
November 14, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27875295/key-residues-and-phosphate-release-routes-in-the-saccharomyces-cerevisiae-pho84-transceptor-the-role-of-tyr179-in-functional-regulation
#20
Dieter R Samyn, Jeroen Van der Veken, Griet Van Zeebroeck, Bengt L Persson, Björn C G Karlsson
Pho84, a major facilitator superfamily (MFS) protein, is the main high-affinity Pi transceptor in Saccharomyces cerevisiae Although transport mechanisms have been suggested for other MFS members, the key residues and molecular events driving transport by Pi:H(+) symporters are unclear. The current Pho84 transport model is based on the inward-facing occluded crystal structure of the Pho84 homologue PiPT in the fungus Piriformospora indica However, this model is limited by the lack of experimental data on the regulatory residues for each stage of the transport cycle...
November 8, 2016: Journal of Biological Chemistry
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