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Protein denaturation

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https://www.readbyqxmd.com/read/28821765/the-protective-action-of-osmolytes-on-the-deleterious-effects-of-gamma-rays-and-atmospheric-pressure-plasma-on-protein-conformational-changes
#1
Pankaj Attri, Minsup Kim, Thapanut Sarinont, Eun Ha Choi, Hyunwoong Seo, Art E Cho, Kazunori Koga, Masaharu Shiratani
Both gamma rays and atmospheric pressure plasma are known to have anticancer properties. While their mechanism actions are still not clear, in some contexts they work in similar manner, while in other contexts they work differently. So to understand these relationships, we have studied Myoglobin protein after the treatment of gamma rays and dielectric barrier discharge (DBD) plasma, and analyzed the changes in thermodynamic properties and changes in the secondary structure of protein after both treatments. The thermodynamic properties were analyzed using chemical and thermal denaturation after both treatments...
August 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28816641/preparation-of-recombinant-human-thymic-stromal-lymphopoietin-protein
#2
Yanlu Zhang, Liehua Wu, Jingai Yu, JianFeng Mei, Yu Yi, JianShu Chen, Guoqing Ying
Human thymic stromal lymphopoietin (hTSLP) protein plays a central role in inflammation. Characterizing properties of hTSLP requires a recombinant overexpression system that produces correctly folded, active hTSLP. In this report, an efficient overexpression system for production of hTSLP was developed. We constructed expression plasmids of the full-length hTslp gene with or without the signal peptide and transformed the plasmids into E. coli. The design of the recombinant proteins included an N-terminal His-tag, which facilitated purification...
August 17, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28813778/the-bactericidal-effect-of-high-temperature-is-an-essential-resistance-mechanism-of-chicken-macrophage-against-brucella-abortus-infection
#3
Lauren Togonon Arayan, Alisha Wehdnesday Bernardo Reyes, Huynh Tan Hop, Huy Tran Xuan, Eun Jin Baek, Wongi Min, Suk Kim
Knowledge of avian host responses to brucellosis is critical to understanding how avian resists this infection however this mechanism is not well established. On the other hand, temperature has a major involvement in the physiology of living organisms and cell death induced by heat is attributed to protein denaturation. This study demonstrates the direct bactericidal effect of a higher temperature (41ºC) to Brucella (B.) abortus that resulted in the gradual reduction of intracellular bacteria that inhibits bacterial growth within avian macrophage HD11 along an increasing period of time...
August 17, 2017: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28812359/continuous-flow-electrophoresis-of-dna-and-proteins-in-a-two-dimensional-capillary-well-sieve
#4
Lian Duan, Zhen Cao, Levent Yobas
Continuous-flow electrophoresis of macromolecules is demonstrated using an integrated capillary-well sieve arranged into a two-dimensional anisotropic array on silicon. The periodic array features thousands of entropic barriers, each resulting from an abrupt interface between a 2-μm-deep well (channel) and a 70-nm capillary. These entropic barriers owing to two-dimensional confinement within the capillaries are vastly steep in relation to those arising from slits featuring one-dimensional confinement. Thus, the sieving mechanisms can sustain relatively large electric field strengths over a relatively small array area...
August 16, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28808343/structure-formation-during-translocon-unassisted-co-translational-membrane-protein-folding
#5
Nicola J Harris, Eamonn Reading, Kenichi Ataka, Lucjan Grzegorzewski, Kalypso Charalambous, Xia Liu, Ramona Schlesinger, Joachim Heberle, Paula J Booth
Correctly folded membrane proteins underlie a plethora of cellular processes, but little is known about how they fold. Knowledge of folding mechanisms centres on reversible folding of chemically denatured membrane proteins. However, this cannot replicate the unidirectional elongation of the protein chain during co-translational folding in the cell, where insertion is assisted by translocase apparatus. We show that a lipid membrane (devoid of translocase components) is sufficient for successful co-translational folding of two bacterial α-helical membrane proteins, DsbB and GlpG...
August 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28807609/cspb-of-an-arctic-bacterium-polaribacter-irgensii-kopri-22228-confers-extraordinary-freeze-tolerance
#6
Youn Hong Jung, Yoo Kyung Lee, Hong Kum Lee, Kyunghee Lee, Hana Im
Freezing temperatures are a major challenge for life at the poles. Decreased membrane fluidity, uninvited secondary structure formation in nucleic acids, and protein cold-denaturation all occur at cold temperatures. Organisms adapted to polar regions possess distinct mechanisms that enable them to survive in extremely cold environments. Among the cold-induced proteins, cold shock protein (Csp) family proteins are the most prominent. A gene coding for a Csp-family protein, cspB, was cloned from an arctic bacterium, Polaribacter irgensii KOPRI 22228, and overexpression of cspB greatly increased the freeze-survival rates of Escherichia coli hosts, to a greater level than any previously reported Csp...
July 31, 2017: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
https://www.readbyqxmd.com/read/28803731/purification-and-characterization-of-the-colicin-a-immunity-protein-in-detergent-micelles
#7
Ane Metola, Ana M Bouchet, Marian Alonso-Mariño, Tammo Diercks, Lena Mäler, Félix M Goñi, Ana R Viguera
The immunity proteins against pore-forming colicins represent a family of integral membrane proteins that reside in the inner membrane of producing cells. Cai, the colicin A immunity protein, was characterized here in detergent micelles by circular dichroism (CD), size exclusion chromatography, chemical cross-linking, nuclear magnetic resonance (NMR) spectroscopy, cysteine accessibility, and colicin A binding in detergent micelles. Bile-salt derivatives induced extensive protein polymerization that precluded further investigation...
August 10, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28803626/putting-the-piezolyte-hypothesis-under-pressure
#8
Christina M Papini, Pranav P Pandharipande, Catherine A Royer, George I Makhatadze
A group of small molecules that stabilize proteins against high hydrostatic pressure has been classified as piezolytes, a subset of stabilizing cosolutes. This distinction would imply that piezolytes counteract the effects of high hydrostatic pressure through effects on the volumetric properties of the protein. The purpose of this study was to determine if cosolutes proposed to be piezolytes have an effect on the volumetric properties of proteins through direct experimental measurements of volume changes upon unfolding of model proteins lysozyme and ribonuclease A, in solutions containing varying cosolute concentrations...
August 10, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28801743/investigation-of-an-optimal-cell-lysis-method-for-the-study-of-the-zinc-metalloproteome-of-histoplasma-capsulatum
#9
Anna M Donnell, Stephanie Lewis, Sami Abraham, Kavitha Subramanian, Julio Landero Figueroa, George S Deepe, Anne P Vonderheide
This work sought to assess optimal extraction conditions in the study of the metalloproteome of the dimorphic fungus Histoplasma capsulatum. One of the body's responses to H. capsulatum infection is sequestration of zinc within host macrophage (MØ), as reported by Vignesh et al. (Immunity 39:697-710, 2013) and Vignesh et al. (PLOS Pathog 9:E1003815, 2013). Thus, metalloproteins containing zinc were of greatest interest as it plays a critical role in survival of the fungus. One challenge in metalloproteomics is the preservation of the native structure of proteins to retain non-covalently bound metals...
August 12, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28800590/assessment-and-optimization-of-theileria-parva-sporozoite-full-length-p67-antigen-expression-in-mammalian-cells
#10
Giulia Tebaldi, Laura B Williams, Andrea E Verna, Francesca Macchi, Valentina Franceschi, Lindsay M Fry, Donald P Knowles, Gaetano Donofrio
Delivery of various forms of recombinant Theileria parva sporozoite antigen (p67) has been shown to elicit antibody responses in cattle capable of providing protection against East Coast fever, the clinical disease caused by T. parva. Previous formulations of full-length and shorter recombinant versions of p67 derived from bacteria, insect, and mammalian cell systems are expressed in non-native and highly unstable forms. The stable expression of full-length recombinant p67 in mammalian cells has never been described and has remained especially elusive...
August 11, 2017: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/28796944/effect-of-freezing-thermal-pasteurization-and-hydrostatic-pressure-on-fractionation-and-folate-recovery-in-egg-yolk
#11
Nassim Naderi, Yves Pouliot, James D House, Alain Doyen
In this study, the impact of pasteurization and freezing of raw material, as performed at a commercial scale, on egg yolk fractionation and folate recovery was assessed. Freezing induced denaturation of the lipoproteins in egg yolk which prevented further fractionation of the yolk. Thermal pasteurization of egg yolk at 61.1ºC for 3.5 min as well as high hydrostatic pressure (HHP) treatment (400 MPa for 5 min) did not change (p<0.05) the composition of egg yolk or yolk fractions after their recovery by centrifugation...
August 10, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28794932/screening-of-neem-extracts-for-microbial-anti-chaperone-activity-by-employing-in-vitro-enzyme-refolding-assay
#12
Jyoti M Patki, Priyanka Shah
Microbial heat shock proteins (Hsps) play an important role in pathogenesis and development of resistance to existing drugs. New compounds that target microbial molecular chaperones have the potential of combating the challenge of anti-microbial resistance. The present study was aimed at assessing the employment of in vitro enzyme refolding assay to detect anti-chaperone activity of Neem (Azadirachta indica) extracts. Protein extracts of thermotolerant Escherichia coli cells were used as a source of Hsps or chaperones...
October 2017: 3 Biotech
https://www.readbyqxmd.com/read/28792701/predictive-value-of-hepatitis-b-core-related-antigen-hbcrag-during-the-natural-history-of-hepatitis-b-virus-infection
#13
Yu Gou, Yanhua Zhao, Chenli Rao, Shu Feng, Tingting Wang, Dongdong Li, Chuanmin Tao
BACKGROUND: The natural history of HBV infection includes immune tolerance (IT), immune clearance (IC), HBeAg-negative inactive/quiescent carrier (ENQ), and HBeAg-negative hepatitis (ENH) phases. As the current biomarkers for discriminating the four phases still have some weaknesses, additional serological indicators are needed. Hepatitis B core-related antigen (HBcrAg) encoded with the precore/core gene contains denatured HBeAg, HBV core antigen (HBcAg), and a 22-KDa precore protein (p22cr) and has been demonstrated to have a close association with the natural history of hepatitis B infection...
July 1, 2017: Clinical Laboratory
https://www.readbyqxmd.com/read/28782566/small-monomeric-and-highly-stable-near-infrared-fluorescent-markers-derived-from-the-thermophilic-phycobiliprotein-apcf2
#14
Wen-Long Ding, Dan Miao, Ya-Nan Hou, Su-Ping Jiang, Bao-Qin Zhao, Ming Zhou, Hugo Scheer, Kai-Hong Zhao
Biliproteins have extended the spectral range of fluorescent proteins into the region of maximal transmission of most tissues and are favorable for multiplexing, but their application presents considerable challenges. Their fluorescence derives from open-chain tetrapyrrole chromophores which often require the introduction of dedicated reductases and lyases. In addition, their fluorescence yield generally decreases with increasing wavelengths and depends strongly on the state of the binding protein. We report fluorescent biliproteins, termed BDFPs, that are derived from the phycobilisome core subunit, ApcF2: this subunit is induced in the thermophilic cyanobacterium, Chroococcidiopsis thermalis, by far-red light and binds phycocyanobilin non-covalently...
August 3, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28782558/exploring-the-denatured-state-ensemble-by-single-molecule-chemo-mechanical-unfolding-the-effect-of-force-temperature-and-urea
#15
Emily Guinn, Susan Marqusee
While it is widely appreciated that the denatured state of a protein is a heterogeneous conformational ensemble, there is still debate over how this ensemble changes with environmental conditions. Here, we use single-molecule chemo-mechanical unfolding, which combines force and urea using the optical tweezers, together with traditional protein unfolding studies to explore how perturbants commonly used to unfold proteins (urea, force and temperature) affect the denatured state ensemble. We compare the urea m-values, which report on the change in solvent accessible surface area for unfolding, to probe the denatured state as a function of force, temperature and urea...
August 3, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28781996/opinion-hazards-faced-by-macromolecules-when-confined-to-thin-aqueous-films
#16
REVIEW
Robert M Glaeser, Bong-Gyoon Han
Samples prepared for single-particle electron cryo-microscopy (cryo-EM) necessarily have a very high surface-to-volume ratio during the short period of time between thinning and vitrification. During this time, there is an obvious risk that macromolecules of interest may adsorb to the air-water interface with a preferred orientation, or that they may even become partially or fully unfolded at the interface. In addition, adsorption of macromolecules to an air-water interface may occur even before thinning. This paper addresses the question whether currently used methods of sample preparation might be improved if one could avoid such interfacial interactions...
2017: Biophysics Reports
https://www.readbyqxmd.com/read/28780460/nanoscale-trifunctional-bovine-hemoglobin-for-fabricating-molecularly-imprinted-polydopamine-via-pickering-emulsions-hydrogels-polymerization
#17
Yanhua Sun, Shian Zhong
In this work, a novel strategy was developed for the selective recognition of protein based on oil-in-water Pickering emulsions-hydrogels polymerization and interfacial imprinting method. For the first time, nanoscale trifunctional bovine hemoglobin (BHb) played three roles simultaneously during the process, including emulsifier, template protein and sacrificial material for further exposing recognition sites. Prior to acting as stabilizers, bovine hemoglobin colloidal particles (BCPs) were firstly prepared by acidification and heat denaturation...
July 29, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28774748/investigation-on-different-chemical-stability-of-mitochondrial-hsp60-and-its-precursor
#18
Caterina Ricci, Rita Carrotta, Giacoma Cinzia Rappa, Maria Rosalia Mangione, Fabio Librizzi, Pier Luigi San Biagio, Heinz Amenitsch, Maria Grazia Ortore, Silvia Vilasi
In the large class of molecules that maintain protein homeostasis, called molecular chaperones, chaperonins constitute a subclass that specifically assist the correct folding of newly synthesized proteins. Among them, Hsp60 is composed of a double heptameric ring structure with a large central cavity where the unfolded protein binds via hydrophobic interactions and is supported, in this function, by the co-chaperonin Hsp10. Hsp60 is typically located in the mitochondria, but in some pathological situations, such as cancers and chronic inflammatory diseases, Hsp60 accumulates in the cytoplasm...
July 26, 2017: Biophysical Chemistry
https://www.readbyqxmd.com/read/28774556/analyzing-bean-extracts-using-time-dependent-sds-trapping-to-quantify-the-kinetic-stability-of-phaseolin-proteins
#19
Jane Thibeault, Jennifer Church, Brian Ortiz-Perez, Samuel Addo, Shakeema Hill, Areeg Khalil, Malaney Young, Ke Xia, Wilfredo Colón
In common beans and lima bean, the storage protein phaseolin is difficult to degrade and SDS-resistant, a sign of kinetic stability. Kinetically stable proteins (KSPs) are characterized by having a high-energy barrier between the native and denatured states that results in very slow unfolding. Such proteins are resistant to proteolytic degradation and detergents, such as SDS. Here the method SDS-Trapping of Proteins (S-TraP) is applied directly on bean extracts to quantify the kinetic stability of phaseolin in lima bean and several common beans, including black bean, navy bean, and small red bean...
July 31, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28771809/role-of-xist-mir-29a-lin28a-pathway-in-denatured-dermis-and-human-skin-fibroblasts-hsfs-after-thermal-injury
#20
Le Guo, Xu Huang, Pengfei Liang, Pihong Zhang, Minghua Zhang, Licheng Ren, Jizhang Zeng, Xv Cui, Xiaoyuan Huang
Denatured dermis is a part of the dermis in deep burn wound and has the ability to restore normal morphology and function. In our previous study, we revealed that miR-29a downregulation in denatured dermis may help burn wound healing in the later phase, and further enhance type I collagen synthesis. LIN28A, a highly-conserved RNA binding protein expressed during embryogenesis, plays roles in development, pluripotency, metabolism, as well as tissue repair in adults. In the present study, we investigated the functional roles of LIN28A in human skin fibroblasts (HSFs) and extracellular matrix (ECM), and the interaction between miR-29a and LIN28A...
August 3, 2017: Journal of Cellular Biochemistry
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