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https://www.readbyqxmd.com/read/28719588/structures-of-carboxylic-acid-reductase-reveal-domain-dynamics-underlying-catalysis
#1
Deepankar Gahloth, Mark S Dunstan, Daniela Quaglia, Evaldas Klumbys, Michael P Lockhart-Cairns, Andrew M Hill, Sasha R Derrington, Nigel S Scrutton, Nicholas J Turner, David Leys
Carboxylic acid reductase (CAR) catalyzes the ATP- and NADPH-dependent reduction of carboxylic acids to the corresponding aldehydes. The enzyme is related to the nonribosomal peptide synthetases, consisting of an adenylation domain fused via a peptidyl carrier protein (PCP) to a reductase termination domain. Crystal structures of the CAR adenylation-PCP didomain demonstrate that large-scale domain motions occur between the adenylation and thiolation states. Crystal structures of the PCP-reductase didomain reveal that phosphopantetheine binding alters the orientation of a key Asp, resulting in a productive orientation of the bound nicotinamide...
July 17, 2017: Nature Chemical Biology
https://www.readbyqxmd.com/read/28716760/properties-of-purified-cyp2r1-in-a-reconstituted-membrane-environment-and-its-25-hydroxylation-of-20-hydroxyvitamin-d3
#2
Chloe Y S Cheng, Tae-Kang Kim, Saowanee Jeayeng, Andrzej T Slominski, Robert C Tuckey
Recent studies indicate that CYP2R1 is the major 25-hydroxylase catalyzing the first step in vitamin D activation. Since the catalytic properties of CYP2R1 have been poorly studied to date and it is a membrane protein, we examined the purified enzyme in a membrane environment. CYP2R1 was expressed in E. coli and purified by nickel affinity- and hydrophobic interaction-chromatography and assayed in a reconstituted membrane system comprising phospholipid vesicles plus purified human NADPH-P450 oxidoreductase...
July 14, 2017: Journal of Steroid Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/28715177/ribosome-mediated-quinary-interactions-modulate-in-cell-protein-activities
#3
Christopher M DeMott, Subhabrata Majumder, David S Burz, Sergey Reverdatto, Alexander Shekhtman
Ribosomes are present inside bacterial cells at micromolar concentrations and occupy up to 20% of the cell volume. Under these conditions even weak quinary interactions between ribosomes and cytosolic proteins can affect protein activity. By using in-cell and in vitro NMR spectroscopy, and biophysical techniques, we show that the enzymes, adenylate kinase and dihydrofolate reductase, and the respective co-enzymes, ATP and NADPH, bind to ribosomes with micromolar affinity, and that this interaction suppresses the enzymatic activities of both enzymes...
July 17, 2017: Biochemistry
https://www.readbyqxmd.com/read/28714670/cross-linking-furan-modified-kisspeptin-10-to-the-kiss-receptor
#4
Willem Vannecke, Christophe Ampe, Marleen Van Troys, Massimiliano Beltramo, Annemieke Madder
Chemical cross-linking is well-established for investigating protein-protein interactions. Traditionally, photo cross-linking is used but is associated with problems of selectivity and UV toxicity in a biological context. We here describe, with live cells and under normal growth conditions, selective cross-linking of a furan-modified peptide ligand to its membrane-presented receptor with zero toxicity, high efficiency, and spatio-specificity. Furan-modified kisspeptin-10 is covalently coupled to its glycosylated membrane receptor, GPR54(KISS1R)...
July 17, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28712268/the-anti-inflammatory-and-antioxidant-properties-of-1-2-decanediol-and-willow-bark-extract-in-lipopolysaccharide-stimulated-keratinocytes
#5
Roberta DI Caprio, Giuseppe Monfrecola, Anna Balato, Nicola Balato, Franco Gasparri, Raffaella Micillo, Serena Lembo
BACKGROUND: The 1,2-decanediol (S-Mal) is an organic compound belonging to the 1,2- alkanediol family, with two hydroxyl groups located on the first and second carbon of the alkane chain, probably responsible for the enhanced anti-bacterial efficacy. The willow bark total extract (W-Mal) has been used since thousands of years as an herbal remedy for its antipyretic, analgesic, anti-inflammatory and anti-microbial activities. S-Mal is used in cosmetic preparations, whether W- Mal can be topically or systemically administered...
July 13, 2017: Giornale Italiano di Dermatologia e Venereologia: Organo Ufficiale, Società Italiana di Dermatologia e Sifilografia
https://www.readbyqxmd.com/read/28712054/alcohol-exposure-promotes-dna-methyltransferase-dnmt3a-upregulation-through-reactive-oxygen-species-dependent-mechanisms
#6
Federico Miozzo, Hélène Arnould, Aurélie de Thonel, Anne-Laure Schang, Délara Sabéran-Djoneidi, Anne Baudry, Benoît Schneider, Valérie Mezger
Abundant evidence has accumulated showing that fetal alcohol exposure broadly modifies DNA methylation profiles in the brain. DNA methyltransferases (DNMTs), the enzymes responsible for DNA methylation, are likely implicated in this process. However, their regulation by ethanol exposure has been poorly addressed. Here, we show that alcohol exposure modulates DNMT protein levels through multiple mechanisms. Using a neural precursor cell line and primary mouse embryonic fibroblasts (MEFs), we found that ethanol exposure augments the levels of Dnmt3a, Dnmt3b, and Dnmt3l transcripts...
July 15, 2017: Cell Stress & Chaperones
https://www.readbyqxmd.com/read/28711923/acute-myeloid-leukemia-cells-require-6-phosphogluconate-dehydrogenase-for-cell-growth-and-nadph-dependent-metabolic-reprogramming
#7
Haymanti Bhanot, Ellen L Weisberg, Mamatha M Reddy, Atsushi Nonami, Donna Neuberg, Richard M Stone, Klaus Podar, Ravi Salgia, James D Griffin, Martin Sattler
Acute myeloid leukemia (AML) cells are highly dependent on glycolytic pathways to generate metabolic energy and support cell growth, hinting at specific, targetable vulnerabilities as potential novel targets for drug development. Elevated levels of NADPH, a central metabolic factor involved in redox reactions, are common in myeloid leukemia cells, but the significance or biochemical basis underlying this increase is unknown. Using a small molecule analog that efficiently inhibits NADPH-producing enzymes, we found that AML cells require NADPH homeostasis for cell growth...
June 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28710857/ros-mediated-selection-for-increased-nadph-availability-in-escherichia-coli
#8
REVIEW
T Steele Reynolds, Colleen M Courtney, Keesha E Erickson, Lisa M Wolfe, Anushree Chatterjee, Prashant Nagpal, Ryan T Gill
The economical production of chemicals and fuels by microbial processes remains an intense area of interest in biotechnology. A key limitation in such efforts concerns the availability of key co-factors, in this case NADPH, required for target pathways. Many of the strategies pursued for increasing NADPH availability in Escherichia coli involve manipulations to the central metabolism, which can create redox imbalances and overall growth defects. In this study we used a reactive oxygen species based selection to search for novel methods of increasing NADPH availability...
July 15, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28710633/laboratory-scale-hydroxylation-of-steroids-by-p450bm3-variants
#9
Sabrina Hoebenreich, Martin Spinck, Nathalie Nett
This chapter describes the asymmetric hydroxylation of steroids on laboratory preparative scale, using engineered variants of P450BM3 (CYP102A1) as enzyme catalyst. The following protocol covers the creation of an Escherichia coli BL21-Gold (DE3) expression strain, including necessary control experiments like plasmid preparation, test expression, and creation of storage cultures, to verify successful experimental access to recombinant expressed P450BM3 variants. The recombinant expressed P450BM3 variants are obtained as cleared cell lysate and used in a biotransformation setup to hydroxylate 2...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28710558/pathway-engineering-for-the-production-of-%C3%AE-amyrin-and-cycloartenol-in-escherichia-coli-a-method-to-biosynthesize-plant-derived-triterpene-skeletons-in-e-coli
#10
Miho Takemura, Rie Tanaka, Norihiko Misawa
Cycloartenol is biosynthetically the first sterol skeleton, which is metabolized to phytosterols such as β-sitosterol and stigmasterol. β-Amyrin is the most commonly occurring aglycone skeleton for oleanane-type saponins such as glycyrrhizin and saikosaponins. It has been regarded that these cyclic triterpenes are unable to be produced in Escherichia coli, while no reports are available on their production with E. coli. Here, we describe a method to synthesize triterpene skeletons from higher plants, including cycloartenol and β-amyrin...
July 15, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28709950/characterization-of-potent-and-selective-iodonium-class-inhibitors-of-nadph-oxidases
#11
Jiamo Lu, Prabhakar Risbood, Charles T Kane, Md Tafazzal Hossain, Larry Anderson, Kimberly Hill, Anne Monks, Yongzhong Wu, Smitha Antony, Agnes Juhasz, Han Liu, Guojian Jiang, Erik Harris, Krishnendu Roy, Jennifer L Meitzler, Mariam Konaté, James H Doroshow
The NADPH oxidases (NOXs) play a recognized role in the development and progression of inflammation-associated disorders, as well as cancer. To date, several NOX inhibitors have been developed, through either high throughput screening or targeted disruption of NOX interaction partners, although only a few have reached clinical trials. To improve the efficacy and bioavailability of the iodonium class NOX inhibitor diphenylene iodonium (DPI), we synthesized 36 analogs of DPI, focusing on improved solubility and functionalization...
July 11, 2017: Biochemical Pharmacology
https://www.readbyqxmd.com/read/28707553/ziram-inhibits-rat-neurosteroidogenic-5%C3%AE-reductase-1-and-3%C3%AE-hydroxysteroid-dehydrogenase
#12
Ying Su, Huitao Li, Xiaomin Chen, Yiyan Wang, Xiaoheng Li, Jianliang Sun, Ren-Shan Ge
The neurotoxicity of ziram is largely unknown. In this study, we investigated the direct inhibitions of ziram on rat neurosteroid synthetic and metabolizing enzymes, 5α-reductase 1 (SRD5A1), 3α-hydroxysteroid dehydrogenase (AKR1C14), and retinol dehydrogenase 2 (RDH2). Rat SRD5A1, AKR1C14, and RDH2 were cloned and transiently expressed in COS1 cells, and the effects of ziram on these enzymes were measured. Ziram inhibited rat SRD5A1 and AKR1C14 with IC50 values of 1.556 ± 0.078 and 1.017 ± 0.072 μM, respectively, when 1000 nM steroid substrates were used...
July 14, 2017: Toxicology Mechanisms and Methods
https://www.readbyqxmd.com/read/28705933/two-functionally-distinct-nadp-dependent-ferredoxin-oxidoreductases-maintain-the-primary-redox-balance-of-pyrococcus-furiosus
#13
Diep M N Nguyen, Gerrit J Schut, Oleg A Zadvornyy, Monika Tokmina-Lukaszewska, Saroj Poudel, Gina L Lipscomb, Leslie A Adams, Jessica T Dinsmore, William J Nixon, Eric S Boyd, Brian Bothner, John W Peters, Michael W W Adams
Electron bifurcation has recently gained acceptance as the third mechanism of energy conservation in which energy is conserved through the coupling of exergonic and endergonic reactions. A structure-based mechanism of bifurcation has been elucidated recently for the flavin-based enzyme NADH-dependent ferredoxin NADP+ oxidoreductase I (NfnI) from the hyperthermophillic archaeon Pyrococcus furiosus. NfnI is thought to be involved in maintaining the cellular redox balance, producing NADPH for biosynthesis by recycling the two other primary redox carriers, NADH and ferredoxin...
July 13, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28705010/isocitrate-dehydrogenase-mutation-as-a-therapeutic-target-in-gliomas
#14
Catherine H Han, Tracy T Batchelor
Isocitrate dehydrogenases (IDH) are important enzymes that catalyze the oxidative decarboxylation of isocitrate to α-ketoglutarate (α-KG), producing NADPH in the process. More than 80% of low-grade gliomas and secondary glioblastoma (GBM) harbor an IDH mutation. IDH mutations involve the catalytic pocket of the enzyme and lead to a neomorphic ability to produce 2-hydroxyglutarate (2HG) while oxidizing NADPH to NADP+. 2HG is considered as an 'oncometabolite' which is thought to be responsible for many, if not all, biologic effects of IDH mutations...
June 2017: Chinese Clinical Oncology
https://www.readbyqxmd.com/read/28704938/nadph-oxidases-angiogenesis-and-peripheral-artery-disease
#15
REVIEW
Pradeep Manuneedhi Cholan, Siân P Cartland, Mary M Kavurma
Peripheral artery disease (PAD) is caused by narrowing of arteries in the limbs, normally occurring in the lower extremities, with severe cases resulting in amputation of the foot or leg. A potential approach for treatment is to stimulate the formation of new blood vessels to restore blood flow to limb tissues. This is a process called angiogenesis and involves the proliferation, migration, and differentiation of endothelial cells. Angiogenesis can be stimulated by reactive oxygen species (ROS), with NADPH oxidases (NOX) being a major source of ROS in endothelial cells...
July 12, 2017: Antioxidants (Basel, Switzerland)
https://www.readbyqxmd.com/read/28704834/gper-mediates-functional-endothelial-aging-in-renal-arteries
#16
Matthias R Meyer, Thomas Rosemann, Matthias Barton, Eric R Prossnitz
Aging is associated with impaired renal artery function, which is partly characterized by arterial stiffening and a reduced vasodilatory capacity due to excessive generation of reactive oxygen species by NADPH oxidases (Nox). The abundance and activity of Nox depends on basal activity of the heptahelical transmembrane receptor GPER; however, whether GPER contributes to age-dependent functional changes in renal arteries is unknown. This study investigated the effect of aging and Nox activity on renal artery tone in wild-type and GPER-deficient (Gper-/-) mice (4 and 24 months old)...
July 14, 2017: Pharmacology
https://www.readbyqxmd.com/read/28704703/early-axis-growth-during-seed-germination-is-gravitropic-and-mediated-by-ros-and-calcium
#17
Khangembam Lenin Singh, Anindita Mukherjee, Rup Kumar Kar
In plant establishment, seed germination is characterized by the emergence of a radicle for secured anchorage to the soil and nutrient and water uptake. Early growth of germinating axes appears to be gravisensitive, and the regulation of this process is largely uncharacterized, particularly in case of epigeally germinating species. Our previous work on the germination of Vigna radiata seeds demonstrated the role of apoplastic reactive oxygen species (ROS) in germination-associated axis growth. This study attempts to explore a possibly similar role of ROS in the gravitropic bending of germinating axes...
July 4, 2017: Journal of Plant Physiology
https://www.readbyqxmd.com/read/28704617/resveratrol-prevents-angiotensin-ii-induced-hypertrophy-of-vascular-smooth-muscle-cells-through-the-transactivation-of-growth-factor-receptors
#18
Ekhtear Hossain, Madhu B Anand-Srivastava
We previously showed that the augmented levels of endogenous angiotensin II (Ang II) contribute to vascular smooth muscle cell (VSMC) hypertrophy through the transactivation of growth factor receptors in spontaneously hypertensive rats. Resveratrol (RV), a polyphenolic component of red wine, has also been shown to attenuate Ang II-evoked VSMC hypertrophy, however, the molecular mechanism mediating this response is obscure. The present study is therefore undertaken to examine if RV could prevent Ang II-induced VSMC hypertrophy through the transactivation of growth factor receptor and associated signaling pathways...
July 13, 2017: Canadian Journal of Physiology and Pharmacology
https://www.readbyqxmd.com/read/28703274/pterostilbene-reduces-oxidative-stress-prevents-hypertrophy-and-preserves-systolic-function-of-right-ventricle-in-cor-pulmonale-model
#19
Denise Dos Santos Lacerda, Patrick Türck, Bruna Gazzi de Lima-Seolin, Rafael Colombo, Vanessa Duarte Ortiz, Jéssica Hellen Poletto Bonetto, Cristina Campos-Carraro, Sara Elis Bianchi, Adriane Belló-Klein, Valquiria LinckBassani, Alex Sander da Rosa Araujo
BACKGROUND AND PURPOSE: In cor pulmonale, the increased afterload imposed on the right ventricle (RV) generates a maladaptive response, impairing the contractile cardiac function. Oxidative mechanisms play an important role in the pathophysiology and progression of this disease. The administration of pterostilbene (PTS), a phyto-phenol with antioxidant potential, may represent a therapeutic alternative. In the present study, the objective was to evaluate the effect of PTS complexed with hydroxypropyl-β-cyclodextrin (HPβCD) on hypertrophy, contractile function and oxidative parameters in the RV of rats with pulmonary hypertension induced by monocrotaline administration...
July 12, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28702667/microengineered-cultures-containing-human-hepatic-stellate-cells-and-hepatocytes-for-drug-development
#20
Matthew D Davidson, David A Kukla, Salman R Khetani
In non-alcoholic steatohepatitis (NASH), hepatic stellate cells (HSC) differentiate into myofibroblast-like cells that cause fibrosis, which predisposes patients to cirrhosis and hepatocellular carcinoma. Thus, modeling interactions between activated HSCs and hepatocytes in vitro can aid in the development of anti-NASH/fibrosis therapeutics and lead to a better understanding of disease progression. Species-specific differences in drug metabolism and disease pathways now necessitate the supplementation of animal studies with data acquired using human liver models; however, current models do not adequately model the negative effects of primary human activated HSCs on the phenotype of otherwise well-differentiated primary human hepatocytes (PHHs) as in vivo...
July 12, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
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