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Protein denaturation

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https://www.readbyqxmd.com/read/28534820/antioxidant-properties-of-probiotic-bacteria
#1
REVIEW
Yang Wang, Yanping Wu, Yuanyuan Wang, Han Xu, Xiaoqiang Mei, Dongyou Yu, Yibing Wang, Weifen Li
Oxidative stress defines a condition in which the prooxidant-antioxidant balance in the cell is disturbed, resulting in DNA hydroxylation, protein denaturation, lipid peroxidation, and apoptosis, ultimately compromising cells' viability. Probiotics have been known for many beneficial health effects, and the consumption of probiotics alone or in food shows that strain-specific probiotics can present antioxidant activity and reduce damages caused by oxidation. However, the oxidation-resistant ability of probiotics, especially the underling mechanisms, is not properly understood...
May 19, 2017: Nutrients
https://www.readbyqxmd.com/read/28534560/re-configurable-multi-mode-contained-electrospray-ionization-for-protein-folding-and-unfolding-on-the-millisecond-time-scale
#2
Colbert F Miller, Dmytro S Kulyk, Jongin W Kim, Abraham K Badu-Tawiah
A new reconfigurable contained-electrospray (ES) ion source is described that can be operated in three unique modes (Types I, II and III) and was applied to control the charge state of proteins for subsequent online characterization by mass spectrometry. Using this device, proteins prepared in 100% water were highly charged after exposure to hydrochloric acid vapor. For myoglobin, the shift to a higher charge state occurred faster than the heme cofactor could escape the unfolding protein. This effect reflected in the detection of highly charged holo-myoglobin intermediates (+26), which suggests the modification process occurred on a millisecond time scale (Type I mode)...
May 23, 2017: Analyst
https://www.readbyqxmd.com/read/28533388/selective-lowering-of-synapsins-induced-by-oligomeric-%C3%AE-synuclein-exacerbates-memory-deficits
#3
Megan E Larson, Susan J Greimel, Fatou Amar, Michael LaCroix, Gabriel Boyle, Mathew A Sherman, Hallie Schley, Camille Miel, Julie A Schneider, Rakez Kayed, Fabio Benfenati, Michael K Lee, David A Bennett, Sylvain E Lesné
Mounting evidence indicates that soluble oligomeric forms of amyloid proteins linked to neurodegenerative disorders, such as amyloid-β (Aβ), tau, or α-synuclein (αSyn) might be the major deleterious species for neuronal function in these diseases. Here, we found an abnormal accumulation of oligomeric αSyn species in AD brains by custom ELISA, size-exclusion chromatography, and nondenaturing/denaturing immunoblotting techniques. Importantly, the abundance of αSyn oligomers in human brain tissue correlated with cognitive impairment and reductions in synapsin expression...
May 22, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28531193/raman-microscopy-investigation-of-vitrification-induced-structural-damages-in-mature-bovine-oocytes
#4
Giulia Rusciano, Carolina De Canditiis, Gianluigi Zito, Marcello Rubessa, Maria Serena Roca, Rosa Carotenuto, Antonio Sasso, Bianca Gasparrini
Although oocyte cryopreservation has great potentials in the field of reproductive technologies, it still is an open challenge in the majority of domestic animals and little is known on the biochemical transformation induced by this process in the different cellular compartments. Raman micro-spectroscopy allows the non-invasive evaluation of the molecular composition of cells, based on the inelastic scattering of laser photons by vibrating molecules. The aim of this work was to assess the biochemical modifications of both the zona pellucida and cytoplasm of vitrified/warmed in vitro matured bovine oocytes at different post-warming times...
2017: PloS One
https://www.readbyqxmd.com/read/28531112/effects-of-polyphenols-on-oxidative-stress-mediated-injury-in-cardiomyocytes
#5
REVIEW
Rosanna Mattera, Monica Benvenuto, Maria Gabriella Giganti, Ilaria Tresoldi, Francesca Romana Pluchinotta, Sonia Bergante, Guido Tettamanti, Laura Masuelli, Vittorio Manzari, Andrea Modesti, Roberto Bei
Cardiovascular diseases are the main cause of mortality and morbidity in the world. Hypertension, ischemia/reperfusion, diabetes and anti-cancer drugs contribute to heart failure through oxidative and nitrosative stresses which cause cardiomyocytes nuclear and mitochondrial DNA damage, denaturation of intracellular proteins, lipid peroxidation and inflammation. Oxidative or nitrosative stress-mediated injury lead to cardiomyocytes apoptosis or necrosis. The reactive oxygen (ROS) and nitrogen species (RNS) concentration is dependent on their production and on the expression and activity of anti-oxidant enzymes...
May 20, 2017: Nutrients
https://www.readbyqxmd.com/read/28530298/aptabody-aptatope-interactions-in-aptablotting-assays
#6
Simranjeet Singh Sekhon, Hyun-Ju Um, Woo-Ri Shin, Sang-Hee Lee, Jiho Min, Ji-Young Ahn, Yang-Hoon Kim
We demonstrate an aptablotting assay method that involves direct and indirect aptabody recognition. Nanoscale single-stranded DNA aptamers against GST and DIG-tags are utilized as aptabodies (GST-2 and DIG-1, respectively), and the GST-2 aptabody binding site, or aptatope, as predicted by a MOE-docking simulation of the protein-aptamer complex, shows the interaction of the GST-2 aptabody at the catalytically active region. The aptabody-aptatope interaction was evaluated by an in vitro enzyme inhibitory analysis...
May 22, 2017: Nanoscale
https://www.readbyqxmd.com/read/28528950/effects-of-cataract-causing-mutations-w59c-and-w151c-on-%C3%AE-b2-crystallin-structure-stability-and-folding
#7
Wei-Jie Zhao, Jia Xu, Xiang-Jun Chen, Hui-Hui Liu, Ke Yao, Yong-Bin Yan
β/γ-Crystallins, the predominant structural proteins in vertebrate lens with lifelong stability to maintain lens transparency, share a high similarity in their primary sequences and tertiary structures. Four conserved Trp residues have been shown to be important to γ-crystallin structure, stability and protection against UV irradiation, whereas their roles in β-crystallins remain elusive. Herein we found that two congenital cataract-causing mutations, W59C and W151C, dramatically decreased βB2-crystallin solubility and stability against thermal and guanidine hydrochloride-induced denaturation...
May 18, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28528000/biophysical-insight-into-structure-function-relation-of-allium-sativum-protease-inhibitor-by-thermal-chemical-and-ph-induced-modulation-using-comprehensive-spectroscopic-analysis
#8
Tooba Naz Shamsi, Romana Parveen, Huma Naz, Md Anzarul Haque, Sadaf Fatima
In this study, we have analyzed the structural and functional changes in the nature of Allium sativum Protease Inhibitor (ASPI) on undergoing various denaturation with variable range of pH, temperature and urea (at pH 8.2). ASPI being anti-tryptic in nature has native molecular mass of ∼15kDa. The conformational stability, functional parameters and their correlation were estimated under different conditions using circular dichroism, fluorescence and activity measurements. ASPI was found to fall in belongs to α+β protein...
May 17, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28525394/modified-denatured-lysozyme-effectively-solubilizes-fullerene-c60-nanoparticles-in-water
#9
Marialuisa Siepi, Jane Politi, Principia Dardano, Angela Amoresano, Luca De Stefano, Daria Maria Monti, Eugenio Notomista
Fullerenes, allotropic forms of carbon, have very interesting pharmacological effects and engineering applications. However, a very low solubility both in organic solvents and water hinders their use. Fullerene C60, the most studied among fullerenes, can be dissolved in water only in the form of nanoparticles of variable dimensions and limited stability. Here the effect on the production of C60 nanoparticles by native and denatured hen egg white lysozyme, a highly basic protein, has been systematically studied...
May 19, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28525284/intrinsically-disordered-proteins-as-important-players-during-desiccation-stress-of-the-soybean-radicles
#10
Yun Liu, Jiahui Wu, Nan Sun, Chengjian Tu, Xiaoying Shi, Hua Cheng, Simu Liu, Shuiming Li, Yong Wang, Yizhi Zheng, Vladimir N Uversky
Intrinsically disordered proteins (IDPs) play a variety of important physiological roles in all living organisms. However, there is no comprehensive analysis of the abundance of IDPs associated with environmental stress in plants. Here, we show that a set of heat-stable proteins (i.e., proteins that do not denature after boiling at 100°C for 10 min) was present in R0mm and R15mm radicles (i.e., before the radicle emergence and the 15 mm long radicles) of soybean (Glycine max) seeds. This set of 795 iTRAQ-quantified heat-stable proteins contained a high proportion of wholly or highly disordered proteins (15%), which was significantly higher than that estimated for the whole soybean proteome containing 55,787 proteins (9%)...
May 19, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28521505/orientation-and-conformation-of-osteocalcin-adsorbed-onto-calcium-phosphate-and-silica-surfaces
#11
Luisa A Scudeller, Selvi Srinivasan, Alexandre M Rossi, Patrick S Stayton, Gary P Drobny, David G Castner
Adsorption isotherms, circular dichroism (CD) spectroscopy, x-ray photoelectron spectroscopy (XPS), and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were used to investigate the adsorption of human osteocalcin (hOC) and decarboxylated (i.e., Gla converted back to Glu) hOC (dhOC) onto various calcium phosphate surfaces as well as silica surfaces. The adsorption isotherms and XPS nitrogen signals were used to track the amount of adsorbed hOC and dhOC. The intensities of key ToF-SIMS amino acid fragments were used to assess changes in the structure of adsorbed hOC and dhOC...
May 18, 2017: Biointerphases
https://www.readbyqxmd.com/read/28520399/an-immunofluorescence-imaging-strategy-for-evaluation-of-the-accessibility-of-dna-5-hydroxymethylcytosine-in-chromatins
#12
Shangwei Zhong, Zhe Li, Ting Jiang, Xiangjun Li, Hailin Wang
DNA 5-hydroxymethylcytosine (5hmC) is an important epigenetic modification found in various mammalian cells. Immunofluorescence imaging analysis essentially provides visual pictures to the abundance and distribution of DNA 5hmC in single cells. However, nuclear DNA is usually wrapped around nucleosomes and packaged into chromatins, and further bound with many functional proteins. These physiologically relevant events would generate barriers to the anti-5hmC antibody to selectively recognize 5hmC in DNA. By taking advantage of these naturally generated barriers, here we present a strategy to evaluate the accessibility of DNA 5hmC in chromatins in situ...
May 18, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28518087/multimer-page-a-method-for-capturing-and-resolving-protein-complexes-in-biological-samples
#13
Tyler Rhinesmith, Bryan A Killinger, Akhil Sharma, Anna Moszczynska
There are many well-developed methods for purifying and studying single proteins and peptides. However, most cellular functions are carried out by networks of interacting protein complexes, which are often difficult to investigate because their binding is non-covalent and easily perturbed by purification techniques. This work describes a method of stabilizing and separating native protein complexes from unmodified tissue using two-dimensional polyacrylamide gel electrophoresis. Tissue lysate is loaded onto a non-denaturing blue-native polyacrylamide gel, then an electric current is applied until the protein migrates a short distance into the gel...
May 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28516908/an-optimized-strategy-to-measure-protein-stability-highlights-differences-between-cold-and-hot-unfolded-states
#14
Caterina Alfano, Domenico Sanfelice, Stephen R Martin, Annalisa Pastore, Piero Andrea Temussi
Macromolecular crowding ought to stabilize folded forms of proteins, through an excluded volume effect. This explanation has been questioned and observed effects attributed to weak interactions with other cell components. Here we show conclusively that protein stability is affected by volume exclusion and that the effect is more pronounced when the crowder's size is closer to that of the protein under study. Accurate evaluation of the volume exclusion effect is made possible by the choice of yeast frataxin, a protein that undergoes cold denaturation above zero degrees, because the unfolded form at low temperature is more expanded than the corresponding one at high temperature...
May 18, 2017: Nature Communications
https://www.readbyqxmd.com/read/28516207/enhancing-h2o2-resistance-of-an-esterase-from-pyrobaculum-calidifontis-by-structure-guided-engineering-of-the-substrate-binding-site
#15
Pengfei Zhou, Dongming Lan, Grzegorz Maria Popowicz, Xuping Wang, Bo Yang, Yonghua Wang
Green technologies are attracting increasing attention in industrial chemistry where enzymatic reactions can replace dangerous and environmentally unfriendly chemical processes. In situ enzymatic synthesis of peroxycarboxylic acid is an attractive alternative for several industrial applications although concentrated H2O2 can denature the biocatalyst, limiting its usefulness. Herein, we report the structure-guided engineering of the Pyrobaculum calidifontis esterase (PestE) substrate binding site to increase its stability and perhydrolysis activity...
May 17, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28515404/insolubility-and-alteration-of-allergenic-activity-of-wheat-proteins-in-processed-foods
#16
Kajiyo Tanaka, Yuuki Kanie, Michita Naitou, Misa Suzuki, Harue Umemura, Kazunori Tagami, Kazunori Sakai, Tomoko Furuta, Chikako Yamada, Hidehiko Izumi, Tomoharu Yokooji, Hiroaki Matsuo, Komei Ito
BACKGROUND: Food processing causes decomposition, denaturation or polymerization of protein, which may alter an allergic reaction. This study aimed to investigate the insolubility and alteration of wheat allergens in processed foods and the reactivity to patient sera. METHODS: We extracted proteins from wheat flour, udon and bread using different extracts and conducted SDS-polyacrylamide gel electrophoresis. IgE-immunoblotting was also conducted using sera from children with wheat allergy...
2017: Arerugī, [Allergy]
https://www.readbyqxmd.com/read/28513656/restoring-the-biophysical-properties-of-decellularized-patches-through-recellularization
#17
Gigi Chi Ting Au-Yeung, Udi Sarig, Hadar Sarig, Hanumakumar Bogireddi, Tomer Bronshtein, Limor Baruch, Avihai Spizzichino, Jacob Bortman, Boey Yin Chiang Freddy, Marcelle Machluf, Subbu S Venkatraman
Various extracellular matrix (ECM) scaffolds, isolated through decellularization, were suggested as ideal biomimetic materials for 'Functional tissue engineering' (FTE). The decellularization process comprises a compromise between damaging and preserving the ultrastructure and composition of ECM-previously shown to affect cell survival, proliferation, migration, organization, differentiation and maturation. Inversely, the effects of cells on the ECM constructs' biophysical properties, under physiological-like conditions, remain still largely unknown...
May 17, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/28510015/dissociative-mechanism-for-irreversible-thermal-denaturation-of-oligomeric-proteins
#18
REVIEW
Natalia A Chebotareva, Svetlana G Roman, Boris I Kurganov
Protein stability is a fundamental characteristic essential for understanding conformational transformations of the proteins in the cell. When using protein preparations in biotechnology and biomedicine, the problem of protein stability is of great importance. The kinetics of denaturation of oligomeric proteins may have characteristic properties determined by the quaternary structure. The kinetic schemes of denaturation can include the multiple stages of conformational transitions in the protein oligomer and stages of reversible dissociation of the oligomer...
December 2016: Biophysical Reviews
https://www.readbyqxmd.com/read/28506644/affinity-purification-of-car9-tagged-proteins-on-silica-matrices-optimization-of-a-rapid-and-inexpensive-protein-purification-technology
#19
Jessica Soto-Rodríguez, Brandon L Coyle, Ariana Samuelson, Kannan Aravagiri, François Baneyx
Car9, a dodecapeptide identified by cell surface display for its ability to bind to the edge of carbonaceous materials, also binds to silica with high affinity. The interaction can be disrupted with l-lysine or l-arginine, enabling a broad range of technological applications. Previously, we reported that C-terminal Car9 extensions support efficient protein purification on underivatized silica. Here, we show that the Car9 tag is functional and TEV protease-excisable when fused to the N-termini of target proteins, and that it supports affinity purification under denaturing conditions, albeit with reduced yields...
May 12, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28503125/thawing-of-pooled-solvent-detergent-treated-plasma-octaplaslg%C3%A2-validation-studies-using-different-thawing-devices
#20
Andrea Heger, Katharina Pock, Jürgen Römisch
BACKGROUND: The aim of this study was to perform validation of the thawing process for solvent/detergent-treated plasma octaplasLG® using different thawing devices. Optimized settings for temperature and thawing time should be defined based on the results of both temperature measurements and extensive biochemical characterization studies. METHODS: octaplasLG units were thawed using water bath systems (i.e. MB-13A, QuickThaw® DH4), dry tempering systems (i.e. plasmatherm, SAHARA-III), and microwave oven (i...
April 2017: Transfusion Medicine and Hemotherapy
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