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Archives of Biochemistry and Biophysics

Dinesh K Yadav, Neelam Yadav, Sarika Yadav, Shafiul Haque, Narendra Tuteja
Advancements in peptide fusion technologies to maximize the protein production has taken a big leap to fulfill the demands of post-genomics era targeting elucidation of structure/function of the proteome and its therapeutic applications, by over-expression in heterologous expression systems. Despite being most preferred protein expression system armed with variety of cardinal fusion tags, expression of the functionally active recombinant protein in E. coli remains plagued. The present review critically analyses the aptness of well-characterized fusion tags utilized for over-expression of recombinant proteins with improved solubility and their compatibility with downstream purification procedures...
October 19, 2016: Archives of Biochemistry and Biophysics
Virve Sihvola, Anna-Liisa Levonen
Reactive oxygen species (ROS) and products of their reactions with cellular macromolecules such as unsaturated fatty acids have been implicated to be important regulators of signalling processes via oxidation or alkylation of redox active thiol residues in target proteins. One of key redox-sensitive signalling proteins mediating the response to oxidant stress is Keap1 (Kelch-like erythroid cell-derived protein with CNC homology [ECH]-associated protein 1), which is a negative regulator of transcription factor Nuclear factor-erythroid 2 p45-related factor 2 (Nrf2) and the central hub for sensing endogenous and environmental oxidative and electrophilic stress...
October 18, 2016: Archives of Biochemistry and Biophysics
Brett A Beaupre, Joseph V Roman, Matthew R Hoag, Kathleen M Meneely, Nicholas R Silvaggi, Audrey L Lamb, Graham R Moran
Renalase catalyzes the oxidation of isomers of β-NAD(P)H that carry the hydride in the 2 or 6 positions of the nicotinamide base to form β-NAD(P)(+). This activity is thought to alleviate inhibition of multiple β-NAD(P)-dependent enzymes of primary and secondary metabolism by these isomers. Here we present evidence for a variety of ligand binding phenomena relevant to the function of renalase. We offer evidence of the potential for primary metabolism inhibition with structures of malate dehydrogenase and lactate dehydrogenase bound to the 6-dihydroNAD isomer...
October 18, 2016: Archives of Biochemistry and Biophysics
Elizabeth C Ledgerwood, James W A Marshall, Johannes F Weijman
Peroxiredoxin 1 is a member of the ubiquitous peroxiredoxin family of thiol peroxidases that catalyse the reduction of peroxides. In recent years eukaryotic peroxiredoxins have emerged as a critical component of cellular redox signalling, particularly in response to alterations in production of hydrogen peroxide. Peroxiredoxins are exquisitely sensitive to oxidation by hydrogen peroxide making them key peroxide sensing enzymes within cells. Evidence gathered over the last decade suggests that in addition to sensing the redox signal, peroxiredoxins have a major role in transducing this signal to downstream signalling proteins, ultimately contributing to regulation of diverse cellular processes including proliferation, differentiation and apoptosis...
October 15, 2016: Archives of Biochemistry and Biophysics
Nicole M Hjortland, Andrew D Mesecar
USP17 is a deubiquitinating enzyme that is upregulated in numerous cancers and therefore a drug target. We developed a robust expression, purification, and assay system for USP17 enabling its enzymatic and structural characterization. USP17 was expressed in E. coli as inclusion bodies and then solubilized, refolded, and purified using affinity and size-exclusion chromatography. Milligram quantities of pure USP17 can be produced that is catalytically more efficient (kcat/Km = 1500 (x10(3)) M(-1) sec(-1)) than other human USPs studied to date...
October 15, 2016: Archives of Biochemistry and Biophysics
Monika Łęgowska, Yveline Hamon, Anna Wojtysiak, Renata Grzywa, Marcin Sieńczyk, Timo Burster, Brice Korkmaz, Adam Lesner
Cathepsin C is a wildly expressed cysteine exopeptidase that is mostly recognized for the activation of the granule-associated proinflammatory serine proteases in neutrophils, cytotoxic T lymphocytes and mast cells. It has been shown that the enzyme can be secreted extracellularly; however, its occurrence in human bodily fluids/physiological samples has not been thoroughly studied. In the course of this study, the first fluorescence resonance energy transfer peptides for the measurement of the activity of human cathepsin C were designed and synthesized...
October 13, 2016: Archives of Biochemistry and Biophysics
Yarrow Madrona, Christopher A Waddling, Paul R Ortiz de Montellano
DosS is a sensor in Mycobacterium tuberculosis that differentially responds to O2, NO, and CO, as well as to changes in the redox state of the prosthetic heme iron atom. The ferrous protein and its Fe(II)NO and Fe(II)CO complexes undergo autophosphorylation and subsequently transfer the phosphate group to DosR, a nuclear factor, to activate it. In contrast, autophosphorylation is negligible with the ferric protein and the Fe(II)O2 complex. To clarify the basis for this differential response to gases, we have determined the crystal structures of the NO and COcomplexes of the DosS GAF-A domain, which contains the heme to which the gases bind...
October 8, 2016: Archives of Biochemistry and Biophysics
Yu Gao, Lanyan Zheng, Jian-Jun Li, Yuguang Du
Two structural Ca(2+) (proximal and distal) is known to be important for ligninolytic peroxidases. However, few studies toward impact of residues involved in two Ca(2+) on properties of ligninolytic peroxidases have been done, especially the proximal one. In this study, mutants of nine residues involved in liganding two Ca(2+) of Pleurotus eryngii versatile peroxidase (VP) were investigated. Most mutants almost completely lost activities, except the mutants of proximal Ca(2+) - S170A and V192T. In comparison with WT (wild type), optimal pH values of S170A, S170D, and V192T shifted from pH 3...
October 5, 2016: Archives of Biochemistry and Biophysics
Andrés Trostchansky, Homero Rubbo
Nitrated derivatives of unsaturated fatty acids (nitro-fatty acids) are being formed and detected in human plasma, cell membranes and tissue, triggering signaling cascades via covalent and reversible post-translational modifications of nucleophilic amino acids in transcriptional regulatory proteins. Arachidonic acid (AA) represents a precursor of potent signaling molecules, i.e., prostaglandins and thromboxanes through enzymatic and non-enzymatic oxidative pathways. Arachidonic acid can be nitrated by reactive nitrogen species leading to the formation of nitro-arachidonic acid (NO2-AA)...
October 5, 2016: Archives of Biochemistry and Biophysics
Michelle Heirbaut, Frederik Lermyte, Esther M Martin, Steven Beelen, Tim Verschueren, Frank Sobott, Sergei V Strelkov, Stephen D Weeks
Small heat shock proteins are ATP-independent molecular chaperones. Their function is to bind partially unfolded proteins under stress conditions. In vivo, members of this chaperone family are known to preferentially assemble together forming large, polydisperse heterooligomers. The exact molecular mechanisms that drive specific heteroassociation are currently unknown. Here we study the oligomers formed between human HSPB1 and HSPB6. Using small-angle X-ray scattering we could characterize two distinct heterooligomeric species present in solution...
October 4, 2016: Archives of Biochemistry and Biophysics
Tarah A Word, Randy W Larsen
Treatment of horse heart Cytochrome-c (Cc) with N-chloro-4-toluosulfonamide (Chloramine-t, CT) results in the oxidation of methionine (Met) residues to the corresponding sulfoxide including the distal heme ligand, Met80. The resulting Fe-sulfoxide coordination is sufficiently labile in the ferrous form to be displaced by gaseous ligands, including CO. Photolysis of the CO-CT-Cc complex provides an opportunity to examine ligand binding dynamics that are associated with a relatively rigid distal heme pocket. In this work, photoacoustic calorimetry (PAC) was utilized to obtain the kinetics as well as enthalpy and molar volume changes subsequent to CO photo-dissociation from CO-CT-Cc...
October 4, 2016: Archives of Biochemistry and Biophysics
Ernesto Cuevasanta, Matías N Möller, Beatriz Alvarez
Hydrogen sulfide (H2S) has been traditionally considered to be a toxic molecule for mammals. However, it can be formed endogenously and exert physiological effects with potential health benefits. H2S can partition two-fold in biological membranes and traverse them rapidly, diffusing between compartments. H2S reactivity has similarities to that of thiols, although it is less nucleophilic than thiols and it can form different products. H2S can react with oxidants derived from the partial reduction of oxygen, but direct scavenging is unlikely to explain H2S protective actions...
September 30, 2016: Archives of Biochemistry and Biophysics
Benjamin Kalbe, Marian Schlimm, Sebastian Wojcik, Stathis Philippou, Désirée Maßberg, Fabian Jansen, Paul Scholz, Hermann Luebbert, Burkhard Ubrig, Sabrina Osterloh, Hanns Hatt
Cells of the renal tubule system are in direct contact with compounds dissolved in the urine, such as short chain fatty acids (SCFA). Murine OR78, a member of the olfactory receptor (OR) family, is involved in SCFA-related regulation of renal blood pressure in mice. It is still unclear whether OR signaling has an impact on human renal physiology. In our study, we showed that OR51E1 and OR11H7, both of which can be activated by the SCFA isovaleric acid, are expressed in the HK-2 human proximal tubule cell line...
September 28, 2016: Archives of Biochemistry and Biophysics
Zeinab Takalloo, Reza H Sajedi, Saman Hosseinkhani, S Mohsen Asghari
Artemin is an abundant thermostable protein in Artemia encysted embryos and considered as a stress protein, as its highly regulated expression is associated with stress resistance. Artemin cDNA was previously isolated and cloned from Artemia urmiana and artemin was found as an efficient molecular chaperone in vitro. Here, co-transformation of E. coli was performed with two expression vectors containing artemin and firefly luciferase for in vivo studies. The time-course of luciferase inactivation at low and elevated temperatures showed that luciferase was rapidly inactivated in control cells, but it was found that luciferase was protected significantly in artemin expressing cells...
September 28, 2016: Archives of Biochemistry and Biophysics
Seiya Arita, Yuta Kinoshita, Kaori Ushida, Atsushi Enomoto, Kyoko Inagaki-Ohara
Obesity increases the risk for gastric cancers. However, the occurrence and mechanisms of precancerous atrophic gastritis induced by high-fat diet (HFD) remain unclear. Here, we show that HFD-associated lipotoxicity induces precancerous lesions that are accompanied by the disruption of organelle homeostasis, tissue integrity, and deregulated expression of stemness genes in the gastric epithelium mediated by leptin receptor (ObR) signaling. Following HFD feeding, ectopic fat accumulated and expression of LAMP2A in lysosome and COX IV in mitochondria increased in the gastric mucosa...
September 28, 2016: Archives of Biochemistry and Biophysics
Henry Jay Forman, Michael J Davies, Anna C Krämer, Giovanni Miotto, Mattia Zaccarin, Hongqiao Zhang, Fulvio Ursini
Oxidation of critical signaling protein cysteines regulated by H2O2 has been considered to involve sulfenic acid (RSOH) formation. RSOH may subsequently form either a sulfenyl amide (RSNHR') with a neighboring amide, or a mixed disulfide (RSSR') with another protein cysteine or glutathione. Previous studies have claimed that RSOH can be detected as an adduct (e.g., with 5,5-dimethylcyclohexane-1,3-dione; dimedone). Here, kinetic data are discussed which indicate that few proteins can form RSOH under physiological signaling conditions...
September 28, 2016: Archives of Biochemistry and Biophysics
Haiting Zhao, Renjun Peng, Qing Liu, Dingyang Liu, Peng Du, Jian Yuan, Gang Peng, Yiwei Liao
H19, one of the first found cancer-associated long non-coding RNAs (lncRNAs), is involved in the development and progression of many types of tumors. An aberrant expression of H19 was observed in hepatocellular carcinoma, cervical cancer, breast cancer, ovarian cancer, and colorectal cancer. However, the exact effects and molecular mechanisms of H19 in glioma progression are still unknown up to now. In this study, we investigated the role of H19 in human glioma cell lines and clinical tumor samples in order to determine the function of this molecule...
September 28, 2016: Archives of Biochemistry and Biophysics
Willem H Koppenol, Patricia L Bounds
There is currently interest in sulfur-containing molecules that may or may not play a role in signaling. We have collected relevant thermodynamic data, namely standard Gibbs energies of formation and electrode potentials at pH 7, and used these to construct a Frost diagram. Thermodynamic data not available in the literature could be estimated with reasonable confidence. At pH 7, the electrode potential of the RSS/RSS(-) couple is +0.68 V, 0.28 V less than that of the RS, H(+)/RSH couple. S2(-) is unstable with respect to HSS(-) and S2...
September 23, 2016: Archives of Biochemistry and Biophysics
Atsushi Matsuzawa
Reactive oxygen species (ROS) are not only cytotoxic products from external and internal environment, but also important mediators of redox signaling. Therefore, thioredoxin (Trx) as an antioxidant maintains the balance of the thiol-related redox status, and also plays pivotal roles in the regulation of redox signaling. Trx senses and responds to environmental oxidative stress and ROS generated by cellular respiration, metabolism, and immune response, and then modulates the redox status, function, and activity of its target signaling proteins...
September 22, 2016: Archives of Biochemistry and Biophysics
Qiongyan Zou, Haijun Wu, Fenfen Fu, Wenjun Yi, Lei Pei, Meirong Zhou
Raf-1 kinase inhibitor protein (RKIP) is a tumor and metastasis suppressor in cancer cells. MicroRNAs (miRNAs) have been suggested to play a vital role in tumor initiation and progression by negatively regulating oncogenes and tumor suppressors. Quite recently, studies have identified some miRNAs operating to promote or suppress tumor invasion or metastasis via regulating metastasis-related genes, providing potential therapeutic targets on anti-metastasis strategy. In this study, we found the expression of RKIP and miR-185 in breast cancer tissues was significantly lower than that of in normal breast tissues...
September 17, 2016: Archives of Biochemistry and Biophysics
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