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Archives of Biochemistry and Biophysics

Maioli E, Daveri E, Maellaro E, Ietta F, Cresti L, Valacchi G
In the past few years, we focused the interest on rottlerin, an old/new natural substance that, over the time, has revealed a number of cellular and molecular targets, all potentially implicated in the fight against cancer. Past and recent literature well demonstrated that rottlerin is an inhibitor of enzymes, transcription factors and signaling molecules that control cancer cell life and death. Although the rottlerin anticancer activity has been mainly ascribed to apoptosis and/or autophagy induction, recent findings unveiled the existence of additional mechanisms of toxicity...
March 12, 2018: Archives of Biochemistry and Biophysics
Jean Paul Ten Klooster, Alexandros Sotiriou, Sjef Boeren, Stefan Vaessen, Jacques Vervoort, Raymond Pieters
Currently, there is a worldwide increase of patients with Type 2 diabetes (T2D). During the progression of healthy obese to T2D status, there is an influx of immune cells, in particular macrophages, into visceral adipose tissue, accompanied by an increase of inflammatory cytokines, such as, IL6, TNFα and Hp. To get a better insight in the underlying mechanisms, we performed a quantitative LCMS analysis on a modified in vitro assay, combining 3T3L1 adipocytes and activated RAW264.7 macrophages, thus mimicking inflamed adipose tissue...
March 8, 2018: Archives of Biochemistry and Biophysics
Minhal Abidi, M Salman Khan, Saheem Ahmad, Tasneem Kausar, Shahid M Nayeem, Sidra Islam, Asif Ali, Khursheed Alam, Moinuddin
Methylglyoxal (MGO), a reactive dicarbonyl metabolite is a potent arginine directed glycating agent which has implications for diabetes-related complications. Dicarbonyl metabolites are produced endogenously and in a state of misbalance, they contribute to cell and tissue dysfunction through protein and DNA modifications causing dicarbonyl stress. MGO is detoxified by glyoxalase 1 (GLO1) system in the cytoplasm. Reactive oxygen species (ROS) are known to aggravate the glycation process. Both the processes are closely linked, and their combined activity is often referred to as "glycoxidation" process...
March 7, 2018: Archives of Biochemistry and Biophysics
K Koruza, B Lafumat, Á Végvári, W Knecht, S Z Fisher
Deuterated proteins and other bio-derived molecules are important for NMR spectroscopy, neutron reflectometry, small angle neutron scattering, and neutron protein crystallography. In the current study we optimized expression media and cell culture conditions to produce high levels of 3 different deuterated human carbonic anhydrases (hCAs). The labeled hCAs were then characterized and tested for deuterium incorporation by mass spectrometry (MS), temperature stability, and propensity to crystallize. The results show that is possible to get very good yields (>10 mg of pure protein per liter of cell culture under deuterated conditions) and that protein solubility is unaffected at the crystallization concentrations tested...
March 7, 2018: Archives of Biochemistry and Biophysics
Nicoleta T Ploscariu, Nienke W M de Jong, Kok P M van Kessel, Jos A G van Strijp, Brian V Geisbrecht
Staphylococcus aureus and related species are highly adapted to their hosts and have evolved numerous strategies to evade the immune system. S. aureus shows resistance to killing following uptake into the phagosome, which suggests that the bacterium evades intracellular killing mechanisms used by neutrophils. We recently discovered an S. aureus protein (SPIN for Staphylococcal Peroxidase INhibitor) that binds to and inhibits myeloperoxidase (MPO), a major player in the oxidative defense of neutrophils. To allow for comparative studies between multiple SPIN sequences, we identified a panel of homologs from species closely related to S...
March 7, 2018: Archives of Biochemistry and Biophysics
Anshul Chaudhary, Vinod Kumar, Prashant K Singh, Pradeep Sharma, Hridoy R Bairagya, Punit Kaur, Sujata Sharma, Shyam S Chauhan, Tej P Singh
Secretory signalling glycoprotein (SPX-40) from mammary gland is highly expressed during involution. This protein is involved in a programmed cell death during tissue remodelling which occurs at the end of lactation. SPX-40 was isolated and purified from buffalo (SPB-40) from the samples obtained during involution. One solution of SPB-40 was made by dissolving it in buffer containing 25 mM Tris-HCl and 50 mM NaCl at pH 8.0. Another solution was made by adding 25% ethanol to the above solution. The biological effects of SPB-40 dissolved in above two solutions were evaluated on MCF-7 breast cancer cell lines...
March 7, 2018: Archives of Biochemistry and Biophysics
Raffaella Soleti, Ramaroson Andriantsitohaina, M Carmen Martinez
Polyphenols are found in plant-derived foods and beverages and display numerous protective effects against cancers, cardiovascular, metabolic and neurodegenerative diseases. Extracellular vesicles (EVs), microparticles, exosomes, and apoptotic bodies, originated by different cell types are emerging as a novel mean of cell-to-cell communication in physiology and pathology and represent a new way to convey fundamental information between cells. Polyphenols can act on signaling pathways that interfere with the biogenesis of EVs...
March 5, 2018: Archives of Biochemistry and Biophysics
Emiri Hotta, Hirokazu Hara, Tetsuro Kamiya, Tetsuo Adachi
Non-thermal atmospheric pressure plasma (NTAPP) has recently emerged as a novel medical therapy for skin wounds. Interleukin-8 (IL-8) is thought to play a critical role in wound healing. NTAPP irradiation has been reported to promote production of IL-8; however, the mechanism is not fully understood. The aim of this study was to elucidate the underlying mechanism of NTAPP-induced IL-8 expression in human keratinocyte HaCaT cells. NTAPP irradiation of HaCaT cells increased IL-8 mRNA expression in an irradiation time-dependent manner...
March 5, 2018: Archives of Biochemistry and Biophysics
Jiaqi Chu, Yuanyuan Fu, Jiecheng Xu, Xueping Zheng, Qianqian Gu, Xia Luo, Qi Dai, Shuxian Zhang, Peiqing Liu, Liang Hong, Min Li
Atg4 is essential for autophagosome formation and Atg8 recycle with the function of processing the precursor and the lipidated Atg8-family proteins. Abnormal autophagic activity is involved in a variety of pathophysiological diseases and ATG4B is of interest as a potential therapeutic target due to its key roles in autophagy process. So ATG4B inhibitors are highly needed. FMK-9a is the most potent inhibitor reported so far. In this study, we confirmed FMK-9a could suppress ATG4B activity in vitro and in cells, with an IC50 of 260 nM...
March 3, 2018: Archives of Biochemistry and Biophysics
Armen O Simonyan, Vladimir V Sirenko, Olga E Karpicheva, Katarzyna Robaszkiewicz, Małgorzata Śliwinska, Joanna Moraczewska, Zoya I Krutetskaya, Yurii S Borovikov
Using the polarized photometry technique we have studied the effects of two amino acid replacements, E240K and R244G, in tropomyosin (Tpm1.1) on the position of Tpm1.1 on troponin-free actin filaments and the spatial arrangement of actin monomers and myosin heads at various mimicked stages of the ATPase cycle in the ghost muscle fibers. E240 and R244 are located in the C-terminal, seventh actin-binding period, in f and b positions of the coiled-coil heptapeptide repeat. Actin, Tpm1.1, and myosin subfragment-1 (S1) were fluorescently labeled: 1...
March 3, 2018: Archives of Biochemistry and Biophysics
Deyamira Matuz-Mares, Alain Hernández-Vázquez, Héctor Riveros-Rosas, Raquel Guinzberg, Tania Quesada-López, Alfonso Cárabez-Trejo, Ofelia Mora, Enrique Piña
The physiological regulation of hepatic glutathione efflux by catecholamines is poorly understood. The purpose of this work was to review the role of adrenergic receptors (AR) on total glutathione (GT ) efflux in rat liver. Two models were used: isolated hepatocytes and perfused livers. In hepatocytes 10 μM adrenaline (Adr), but not isoproterenol (Iso) a β-AR agonist, or phenylephrine (Phe) an α1 -AR agonist, (in a Krebs-Henseleit Buffer (KHB) enriched with Ca2+ and some aminoacids) increased in 13% GT efflux...
February 26, 2018: Archives of Biochemistry and Biophysics
Pilar C Portela, Tomás M Fernandes, Joana M Dantas, Marisa R Ferreira, Carlos A Salgueiro
G. metallireducens bacterium has highly versatile respiratory pathways that provide the microorganism an enormous potential for many biotechnological applications. However, little is known about the structural and functional properties of its electron transfer components. In this work, the periplasmic cytochrome PpcA from G. metallireducens was studied in detail for the first time using complementary biophysical techniques, including UV-visible, CD and NMR spectroscopy. The results obtained showed that PpcA contains three low-spin c-type heme groups with His-His axial coordination, a feature also observed for its homologue in G...
February 24, 2018: Archives of Biochemistry and Biophysics
Albert Galera-Prat, David Pantoja-Uceda, Douglas V Laurents, Mariano Carrión Vázquez
Bacterial cellulases are drawing increased attention as a means to obtain plentiful chemical feedstocks and fuels from renewable lignocellulosic biomass sources. Certain bacteria deploy a large extracellular multi-protein complex, called the cellulosome, to degrade cellulose. Scaffoldin, a key non-catalytic cellulosome component, is a large protein containing a cellulose-specific carbohydrate-binding module and several cohesin modules which bind and organize the hydrolytic enzymes. Despite the importance of the structure and protein/protein interactions of the cohesin module in the cellulosome, its structure in solution has remained unknown to date...
February 24, 2018: Archives of Biochemistry and Biophysics
Mallory E Gillam, Gregory A Hunter, Gloria C Ferreira
Protoporphyrin ferrochelatase catalyzes the insertion of Fe2+ into protoporphyrin IX to form heme. To determine whether a conserved, active site π-helix contributes to the translocation of the metal ion substrate to the ferrochelatase-bound porphyrin substrate, the invariant π-helix glutamates were replaced with amino acids with non-negatively charged side chains, and the kinetic mechanisms of the generated variants were examined. Analysis of yeast wild-type ferrochelatase-, E314Q- and E318Q-catalyzed reactions, under multi- and single-turnover conditions, demonstrated that the mutations of the π-helix glutamates hindered both protoporphyrin metalation and release of the metalated porphyrin, by slowing each step by approximately 30-50%...
February 23, 2018: Archives of Biochemistry and Biophysics
Jinyoung Suh, Do-Hee Kim, Young-Joon Surh
Cancer-associated fibroblasts (CAFs) constitute a major compartment of the tumor microenvironment. CAFs produce a variety of cytokines, growth factors and extracellular matrix proteins, thereby stimulating tumor progression. CAFs are distinct from normal fibroblasts for their overexpression of α-smooth muscle actin. Recent studies suggest that CAFs play an important role in proliferation and migration of cancer cells through cross-talk with them. Resveratrol (trans-3,4'5,-trihydroxystilbene), a phytoalexin present in grapes, has been reported to possess chemopreventive and chemotherapeutic activities...
February 22, 2018: Archives of Biochemistry and Biophysics
Katarína Šuchová, Stanislav Kozmon, Vladimír Puchart, Anna Malovíková, Tine Hoff, Kristian B R Mørkeberg Krogh, Peter Biely
XynA from Erwinia chrysanthemi (EcXyn30A), belonging to glycoside hydrolase family 30 subfamily 8, is specialized for hydrolysis of 4-O-methylglucuronoxylan (GX). Carboxyl group of 4-O-methylglucuronic acid serves as a substrate recognition element interacting ionically with positively charged Arg293 of the enzyme. We determined kinetic parameters of EcXyn30A on GX, its methyl ester (GXE) and 4-O-methylglucoxylan (GXR) and compared them with behavior of the enzyme variant in which Arg293 was replaced by Ala...
February 22, 2018: Archives of Biochemistry and Biophysics
Evan A Wells, Mark A Anderson, Tonya N Zeczycki
The Ca2+ -dependent deamidation and transamidation activities of transglutaminase 2 (TG2) are important to numerous physiological and pathological processes. Herein, we have examined the steady-state kinetics and15 (V/K) kinetic isotope effects (KIEs) for the TG2-catalyzed deamidation and transamidation of N-Benzyloxycarbonyl-l-Glutaminylglycine (Z-Gln-Gly) using putrescine as the acyl acceptor substrate. Kinetic parameters determined from initial velocity plots are consistent with previously proposed mechanisms...
February 22, 2018: Archives of Biochemistry and Biophysics
Govindarajan Prasanna, Pu Jing
In this study, chemical chaperone like function of cyanidin-3-O-glucoside (C3G) was investigated through fluorescence spectroscopy, UV-visible spectroscopy, circular dichroism spectroscopy, confocal microscopy, scanning electron microscopy and molecular docking studies. Early and advanced glycation inhibitory effect was evaluated by fluorescence spectroscopy and agarose gel electrophoresis. Amyloids were investigated based on their propensity to bind Congo Red (CR) and Thioflavin T (ThT) by multiple microscopic approaches...
February 20, 2018: Archives of Biochemistry and Biophysics
Andrea Nicolussi, Markus Auer, Benjamin Sevcnikar, Martina Paumann-Page, Vera Pfanzagl, Marcel Zámocký, Stefan Hofbauer, Paul G Furtmüller, Christian Obinger
Four heme peroxidase superfamilies arose independently in evolution. Only in the peroxidase-cyclooxygenase superfamily the prosthetic group is posttranslationally modified (PTM). As a consequence these peroxidases can form one, two or three covalent bonds between heme substituents and the protein. This may include ester bonds between heme 1- and 5-methyl groups and glutamate and aspartate residues as well as a sulfonium ion link between the heme 2-vinyl substituent and a methionine. Here the phylogeny and physiological roles of representatives of this superfamily, their occurrence in all kingdoms of life, the relevant sequence motifs for definite identification and the available crystal structures are presented...
February 17, 2018: Archives of Biochemistry and Biophysics
John M Robbins, Andreas S Bommarius, Giovanni Gadda
Formate oxidase (FOX) from Aspergillus oryzae is the only GMC member that oxidizes a carbon acid rather than alcohols; thus, its catalytic mechanism may be different from that of other GMC members. We have used pH, solvent viscosity, and deuterium kinetic isotope effects, to investigate the catalytic mechanism of FOX. The enzyme followed a Bi-Bi sequential steady-state kinetic mechanism. The kcat value was pH-independent between pH 2.8 and 6.8, suggesting lack of ionizable groups in kinetic step(s) that limit the overall turnover of the enzyme...
February 16, 2018: Archives of Biochemistry and Biophysics
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