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Sumana Venkat, Dharma Theja Nannapaneni, Caroline Gregory, Qinglei Gan, Matt McIntosh, Chenguang Fan
Reversible lysine acetylation is one of the most widely distributed post-translational modifications; it is involved in a variety of biological processes and can be found in all three domains of life. Acetyltransferases and deacetylases work coordinately to control levels of protein acetylation. In this work, we applied the genetic code expansion strategy to site-specifically incorporate N(ε)-thioacetyl-l-lysine (TAcK) as an analog of N(ε)-acetyl-l-lysine (AcK) into green fluorescent protein and malate dehydrogenase in Escherichia coli...
November 2017: FEBS Open Bio
David Wright, Zaid Altaany, Yumin Bi, Zaccary Alperstein, Patrick O'Donoghue
TrxR1 is a cancer target and essential selenoprotein that defends the cell against reactive oxygen species and regulates cellular signaling and redox pathways. Previous cell-based studies correlated TrxR1 acetylation with modulated cellular reduction activity, yet the function of specific acetylation sites on TrxR1 remain unknown. We produced site-specifically acetylated TrxR1 variants that also contain selenocysteine (Sec). We demonstrated efficient high-fidelity protein synthesis with 22 different amino acids by simultaneous UAG codon reassignment to Nε-acetyl-lysine (acK) and UGA codon recoding to Sec...
November 8, 2017: Antioxidants & Redox Signaling
Guillaume Acke, Sofie Van Damme, Remco W A Havenith, Patrick Bultinck
The zz component of the nucleus independent chemical shift or the NICSzz is commonly used as a quantifier of the (anti)aromatic character of a (sub)system. One of the underlying assumptions is that a position can be found where the "aromatic" ring currents are adequately reflected in the corresponding NICSzz value. However, as the NICSzz is the result of an integration over the entire space, it no longer explicitly contains the information needed to quantify the separate contributions arising from underlying current density patterns...
November 2, 2017: Journal of Computational Chemistry
Yueting Zheng, Qitao Liu, Huanhuan Shen, Guoyu Yang
Reversible N(ε)-lysine (N(ε)-Lys) acetylation is a dynamic post-translational modification. Genetic incorporation of N(ε)-acetyllysine (N(ε)-AcK) into the specific site of a protein is a powerful method for producing recombinant protein with acetylation and studying the functional role of protein acetylation. Because of the universal existence of deacetylase such as CobB in vivo, the acetyl group of N(ε)-AcK may be removed from recombinant protein. So in the process of incorporating acetyl lysine into protein, nicotinamide (NAM), a lysine deacetylase (KDAC) inhibitor, is needed to inhibit the KDAC activity and protect the acetyl group of N(ε)-acetyllysine incorporated from removal in vivo...
October 3, 2017: Protein Expression and Purification
Jung Ho Ahn, Junho Bang, Won Jun Kim, Sang Yup Lee
There has been much effort exerted to reduce one carbon (C1) gas emission to address climate change. As one promising way to more conveniently utilize C1 gas, several technologies have been developed to convert C1 gas into useful chemicals such as formic acid (FA). In this study, systems metabolic engineering was utilized to engineer Mannheimia succiniciproducens to efficiently utilize FA. (13) C isotope analysis of M. succiniciproducens showed that FA could be utilized through formate dehydrogenase (FDH) reaction and/or the reverse reaction of pyruvate formate lyase (PFL)...
December 2017: Biotechnology and Bioengineering
Luciano Babuin, Daniele Scarpa, Allan S Jaffe
PURPOSE OF REVIEW: Reduction of kidney function and heart disease frequently coexisted in the seam patient. The relation between renal and heart function is quite complex and bring out some unresolved questions about diagnosis (mostly related to the biomarkers levels interpretation), pharmacological therapy (mostly related to drugs kinetics and efficacy), and non-pharmacological therapy. RECENT FINDINGS: Patients with kidney dysfunction (KD) are frequently excluded and underrepresented in the large trials...
August 24, 2017: Current Cardiology Reports
Thanh Dang, Cheryl Ingram-Smith
Acetate kinase (ACK; E.C., which catalyzes the interconversion of acetate and acetyl phosphate, is nearly ubiquitous in bacteria but is present only in one genus of archaea and certain eukaryotic microbes. All ACKs utilize ATP/ADP as the phosphoryl donor/acceptor in the respective directions of the reaction (acetate + ATP [Formula: see text] acetyl phosphate + ADP), with the exception of the Entamoeba histolytica ACK (EhACK) which uses pyrophosphate (PPi)/inorganic phosphate (Pi) (acetyl phosphate + Pi  [Formula: see text] acetate + PPi)...
July 19, 2017: Scientific Reports
Jiaqi Lu, Hua Zhang, Xian Chen, Yong Zou, Jiasong Li, Li Wang, Minhao Wu, Jianye Zang, Yang Yu, Wei Zhuang, Qing Xia, Jiangyun Wang
The modulation of protein acetylation network is a promising strategy for life span extension and disease treatment (Sabari et al., 2016; Giblin et al., 2014) [1,2]. A variety of small molecules have been developed to target deacetylases, but extremely few of these molecules are capable of activating the mitochondrial NAD-dependent deacetylase sirtuin-3 (SIRT3) (Gertz and Steegborn, 2016; Scholz et al., 2015) [3,4]. Manganese superoxide dismutase (MnSOD) is the major superoxide scavenger in mitochondria, whose activity is regulated by SIRT3-mediated deacetylation, particularly at the Lys68 site (Chen et al...
July 12, 2017: Free Radical Biology & Medicine
Cecily S Ray, M S Pednekar, P C Gupta, M Bansal-Travers, Ack Quah, G T Fong
BACKGROUND: Social impacts on tobacco use have been reported but not well quantified. This study investigated how strongly the use of smoked and smokeless tobacco may be influenced by other users who are close to the respondents. METHODS: The International Tobacco Control Project (TCP), India, used stratified multistage cluster sampling to survey individuals aged ≥15 years in four areas of India about their tobacco use and that of their close associates. The present study used logistic regression to calculate odds ratios (ORs) for tobacco use for each type of close associate...
September 2016: WHO South-East Asia Journal of Public Health
Jinhuan Zhang, Jun Long
Wireless sensor networks (WSNs) are one of the important supporting technologies of edge computing. In WSNs, reliable communications are essential for most applications due to the unreliability of wireless links. In addition, network lifetime is also an important performance metric and needs to be considered in many WSN studies. In the paper, an energy-aware hybrid Automatic Repeat-reQuest protocol (ARQ) scheme is proposed to ensure energy efficiency under the guarantee of network transmission reliability. In the scheme, the source node sends data packets continuously with the correct window size and it does not need to wait for the acknowledgement (ACK) confirmation for each data packet...
June 12, 2017: Sensors
Wolfram Volkwein, Christopher Maier, Ralph Krafczyk, Kirsten Jung, Jürgen Lassak
The analysis of the function of essential genes in vivo depends on the ability to experimentally modulate levels of their protein products. Current methods to address this are based on transcriptional or post-transcriptional regulation of mRNAs, but approaches based on the exploitation of translation regulation have so far been neglected. Here we describe a toolbox, based on amber suppression in the presence of N(ε)-acetyl-l-lysine (AcK), for translational tuning of protein output. We chose the highly sensitive luminescence system LuxCDABE as a reporter and incorporated a UAG stop codon into the gene for the reductase subunit LuxC...
June 27, 2017: ACS Synthetic Biology
Weimin Xuan, Peter G Schultz
The use of noncanonical amino acids (ncAAs) to control the viability of an organism provides a strategy for the development of conditional "kill switches" for live vaccines or engineered human cells. We report an approach inspired by the posttranslational acetylation/deacetylation of lysine residues, in which a protein encoded by a gene with an in-frame nonsense codon at an essential lysine can be expressed in its native state only upon genetic incorporation of N-ϵ-acetyl-l-Lys (AcK), and subsequent enzymatic deacetylation in the host cell...
July 24, 2017: Angewandte Chemie
George J N Tetley, Helen R Mott, R Neil Cooley, Darerca Owen
Cdc42 is a Rho-family small G protein that has been widely studied for its role in controlling the actin cytoskeleton and plays a part in several potentially oncogenic signaling networks. Similar to most other small G proteins, Cdc42 binds to many downstream effector proteins to elicit its cellular effects. These effector proteins all engage the same face of Cdc42, the conformation of which is governed by the activation state of the G protein. Previously, the importance of individual residues in conferring binding affinity has been explored for residues within Cdc42 for three of its Cdc42/Rac interactive binding (CRIB) effectors, activated Cdc42 kinase (ACK), p21-activated kinase (PAK), and Wiskott-Aldrich syndrome protein (WASP)...
July 7, 2017: Journal of Biological Chemistry
Lu-Lu Wang, Hui-Hui Guo, Shuai Huang, Chen-Lin Feng, Yan-Xing Han, Jian-Dong Jiang
Short-chain fatty acids (SCFAs) of intestine microbial have caught accumulating attention for their beneficial effects on human health. Botanic compounds with low bioavailability such as berberine (BBR) and resveratrol might interact with intestinal microbial ecosystem and promote gut bacteria to produce SCFA, which contribute to their biological effects. In the present study, a comprehensive assay system was built to detect SCFAs production in intestinal bacteria, in which stringent anaerobic culture was applied for in vitro bacterial fermentation, followed by direct-injection GC detection (chemical detection) in combination with real time polymerase chain reaction (RT-PCR, biological detection)...
July 1, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Qing Ye, Quan-Quan Ji, Wei Yan, Fang Yang, En-Duo Wang
Previous proteomic analyses have shown that aminoacyl-tRNA synthetases in many organisms can be modified by acetylation of Lys. In this present study, leucyl-tRNA synthetase and arginyl-tRNA synthetase from Escherichia coli (EcLeuRS and EcArgRS) were overexpressed and purified and found to be acetylated on Lys residues by MS. Gln scanning mutagenesis revealed that Lys(619), Lys(624), and Lys(809) in EcLeuRS and Lys(126) and Lys(408) in EcArgRS might play important roles in enzyme activity. Furthermore, we utilized a novel protein expression system to obtain enzymes harboring acetylated Lys at specific sites and investigated their catalytic activity...
June 23, 2017: Journal of Biological Chemistry
Chao Xue, Mark Sowden, Bradford C Berk
OBJECTIVE: Oxidative stress and inflammation play key roles in the development of pulmonary arterial hypertension (PAH). Cyclophilin A (CypA) is secreted in response to oxidative stress and promotes inflammation and cardiovascular disease. Endothelial cell (EC) dysfunction is an early event in the pathogenesis of PAH. We evaluated the role of extracellular CypA in PAH and compared the effects of acetylated CypA (AcK-CypA, increased by oxidative stress) and CypA on EC dysfunction. APPROACH AND RESULTS: In transgenic mice that express high levels of CypA in EC specifically, a PAH phenotype was observed at 3 months including increased right ventricular systolic pressure, α-smooth muscle actin expression in small arterioles, and CD45-positive cells in the lungs...
June 2017: Arteriosclerosis, Thrombosis, and Vascular Biology
Andreas H Förster, Sebastian Beblawy, Frederik Golitsch, Johannes Gescher
BACKGROUND: This paper describes the metabolic engineering of Escherichia coli for the anaerobic fermentation of glucose to acetoin. Acetoin has well-established applications in industrial food production and was suggested to be a platform chemical for a bio-based economy. However, the biotechnological production is often hampered by the simultaneous formation of several end products in the absence of an electron acceptor. Moreover, typical production strains are often potentially pathogenic...
2017: Biotechnology for Biofuels
M Moriconi, E Acke, D Petrelli, S Preziuso
Streptococcus canis (S. canis), Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) and Streptococcus dysgalactiae subspecies (S. dysgalactiae subspecies) are β-haemolytic Gram positive bacteria infecting animals and humans. S. canis and S. zooepidemicus are considered as two of the major zoonotic species of Streptococcus, while more research is needed on S. dysgalactiae subspecies bacteria. In this work, a multiplex-PCR protocol was tested on strains and clinical samples to detect S. canis, S. dysgalactiae subspecies and S...
February 2017: Comparative Immunology, Microbiology and Infectious Diseases
Jeong Nam Kim, Robert A Burne
In the dental caries pathogen Streptococcus mutans, phosphotransacetylase (Pta) and acetate kinase (Ack) convert pyruvate into acetate with the concomitant generation of ATP. The genes for this pathway are tightly regulated by multiple environmental and intracellular inputs, but the basis for differential expression of the genes for Pta and Ack in S. mutans had not been investigated. Here, we show that inactivation in S. mutans of ccpA or codY reduced the activity of the ackA promoter, whereas a ccpA mutant displayed elevated pta promoter activity...
April 1, 2017: Applied and Environmental Microbiology
Hong Lin, Hongye Su, Peng Shi, Renquan Lu, Zheng-Guang Wu
In this paper, we study the state estimation and optimal control [i.e., linear quadratic Gaussian (LQG) control] problems for networked control systems in which control inputs, observations, and packet acknowledgments (ACKs) are randomly lost. The packet ACK is a signal that is transmitted from the actuator to notice the estimator the occurence of control packet loss. For such systems, we obtain the optimal estimator, which is consisted of exponentially increasing terms. For the solvability of the LQG problem, we come to a conclusion that in general even the optimal LQG control exists, it is impossible and unnecessary to be obtained as its calculation is not only technically difficult but also computationally prohibitive...
September 13, 2016: IEEE Transactions on Cybernetics
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