Branched chain amino Acids as in vitro and in vivo Anti-Oxidation Compounds

2021 ◽  
pp. 3899-3904
Author(s):  
Moath Alqaraleh ◽  
Violet Kasabri ◽  
Ibrahim Al-Majali ◽  
Nihad Al-Othman ◽  
Nihad Al-Othman ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Amino Acids ◽  
Branched Chain Amino Acids ◽  
Raw 264.7 ◽  
Raw 264.7 Cell ◽  
Branched Chain ◽  
Raw 264.7 Cell Line

Background and aims: Branched chain amino acids (BCAAs) can be tightly connected to metabolism syndrome (MetS) which can be counted as a metabolic indicator in the case of insulin resistance (IR). The aim of this study was to assess the potential role of these acids under oxidative stress. Material and Methods: the in vitro antioxidant activity of BCAAs was assessed using free radical 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging assays. For further check, a qRT-PCR technique was madefor detection the extent of alterations in gene expression of antioxidative enzymes (catalase and glutathione peroxidase (Gpx)) in lipopolysaccharides (LPS(-induced macrophages RAW 264.7 cell line. Additionally, BCAAs antioxidant activity was evaluated based on plasma H2O2 levels and xanthine oxidase (XO) activity in prooxidative LPS-treated mice. Results: Different concentrations of BCAAs affected on DPPH radical scavenging activity but to lesser extent than the ascorbic acid. Besides, BCAAs obviously upregulated the gene expression levels of catalases and Gpx in LPS-modulated macrophage RAW 264.7 cell line. In vivo BCAAs significantly minimized the level of plasma H2O2 as well as the activity of XO activity under oxidative stress. Conclusion: our current findings suggest that BCAAs supplementation may potentially serve as a therapeutic target for treatment of oxidative stress occurs with atherosclerosis, IR-diabetes, MetS and tumorigenesis.

scholarly journals Molecular Mechanisms Underlying the Positive Stringent Response of the Bacillus subtilis ilv-leu Operon, Involved in the Biosynthesis of Branched-Chain Amino Acids

2008 ◽  
Vol 190 (18) ◽  
pp. 6134-6147 ◽  
Author(s):  
Shigeo Tojo ◽  
Takenori Satomura ◽  
Kanako Kumamoto ◽  
Kazutake Hirooka ◽  
Yasutaro Fujita
Keyword(s):  
Amino Acids ◽  
Bacillus Subtilis ◽  
Transcription Initiation ◽  
Molecular Mechanisms ◽  
Stringent Response ◽  
Branched Chain Amino Acids ◽  
Base Substitution ◽  
Branched Chain

ABSTRACT Branched-chain amino acids are the most abundant amino acids in proteins. The Bacillus subtilis ilv-leu operon is involved in the biosynthesis of branched-chain amino acids. This operon exhibits a RelA-dependent positive stringent response to amino acid starvation. We investigated this positive stringent response upon lysine starvation as well as decoyinine treatment. Deletion analysis involving various lacZ fusions revealed two molecular mechanisms underlying the positive stringent response of ilv-leu, i.e., CodY-dependent and -independent mechanisms. The former is most likely triggered by the decrease in the in vivo concentration of GTP upon lysine starvation, GTP being a corepressor of the CodY protein. So, the GTP decrease derepressed ilv-leu expression through detachment of the CodY protein from its cis elements upstream of the ilv-leu promoter. By means of base substitution and in vitro transcription analyses, the latter (CodY-independent) mechanism was found to comprise the modulation of the transcription initiation frequency, which likely depends on fluctuation of the in vivo RNA polymerase substrate concentrations after stringent treatment, and to involve at least the base species of adenine at the 5′ end of the ilv-leu transcript. As discussed, this mechanism is presumably distinct from that for B. subtilis rrn operons, which involves changes in the in vivo concentration of the initiating GTP.

scholarly journals Regulation of stringent factor by branched-chain amino acids

2018 ◽  
Vol 115 (25) ◽  
pp. 6446-6451 ◽  
Author(s):  
Mingxu Fang ◽  
Carl E. Bauer
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Rhodobacter Capsulatus ◽  
Stringent Response ◽  
Branched Chain Amino Acids ◽  
Hydrolase Activity ◽  
Act Domain ◽  
Branched Chain

When faced with amino acid starvation, prokaryotic cells induce a stringent response that modulates their physiology. The stringent response is manifested by production of signaling molecules guanosine 5′-diphosphate,3′-diphosphate (ppGpp) and guanosine 5′-triphosphate,3′-diphosphate (pppGpp) that are also called alarmones. In many species, alarmone levels are regulated by a multidomain bifunctional alarmone synthetase/hydrolase called Rel. In this enzyme, there is an ACT domain at the carboxyl region that has an unknown function; however, similar ACT domains are present in other enzymes that have roles in controlling amino acid metabolism. In many cases, these other ACT domains have been shown to allosterically regulate enzyme activity through the binding of amino acids. Here, we show that the ACT domain present in theRhodobacter capsulatusRel alarmone synthetase/hydrolase binds branched-chain amino acids valine and isoleucine. We further show that the binding of these amino acids stimulates alarmone hydrolase activity both in vitro and in vivo. Furthermore, we found that the ACT domain present in Rel proteins from many diverse species also binds branched-chain amino acids. These results indicate that the cellular concentration of amino acids can directly affect Rel alarmone synthetase/hydrolase activity, thus adding another layer of control to current models of cellular control of the stringent response.

scholarly journals Inactivation of the ilvB1 gene in Mycobacterium tuberculosis leads to branched-chain amino acid auxotrophy and attenuation of virulence in mice

Microbiology ◽  
2009 ◽  
Vol 155 (9) ◽  
pp. 2978-2987 ◽  
Author(s):  
Disha Awasthy ◽  
Sheshagiri Gaonkar ◽  
R. K. Shandil ◽  
Reena Yadav ◽  
Sowmya Bharath ◽  
...  
Keyword(s):  
Amino Acids ◽  
Mycobacterium Tuberculosis ◽  
Amino Acid ◽  
Mutant Strain ◽  
Branched Chain Amino Acids ◽  
Branched Chain Amino Acid ◽  
Branched Chain ◽  
Kinetics Of

Acetohydroxyacid synthase (AHAS) is the first enzyme in the branched-chain amino acid biosynthesis pathway in bacteria. Bioinformatics analysis revealed that the Mycobacterium tuberculosis genome contains four genes (ilvB1, ilvB2, ilvG and ilvX) coding for the large catalytic subunit of AHAS, whereas only one gene (ilvN or ilvH) coding for the smaller regulatory subunit of this enzyme was found. In order to understand the physiological role of AHAS in survival of the organism in vitro and in vivo, we inactivated the ilvB1 gene of M. tuberculosis. The mutant strain was found to be auxotrophic for all of the three branched-chain amino acids (isoleucine, leucine and valine), when grown with either C6 or C2 carbon sources, suggesting that the ilvB1 gene product is the major AHAS in M. tuberculosis. Depletion of these branched chain amino acids in the medium led to loss of viability of the ΔilvB1 strain in vitro, resulting in a 4-log reduction in colony-forming units after 10 days. Survival kinetics of the mutant strain cultured in macrophages maintained with sub-optimal concentrations of the branched-chain amino acids did not show any loss of viability, indicating either that the intracellular environment was rich in these amino acids or that the other AHAS catalytic subunits were functional under these conditions. Furthermore, the growth kinetics of the ΔilvB1 strain in mice indicated that although this mutant strain showed defective growth in vivo, it could persist in the infected mice for a long time, and therefore could be a potential vaccine candidate.

scholarly journals Genetic and Biochemical Analysis of the Interaction of Bacillus subtilis CodY with Branched-Chain Amino Acids

2009 ◽  
Vol 191 (22) ◽  
pp. 6865-6876 ◽  
Author(s):  
Anuradha C. Villapakkam ◽  
Luke D. Handke ◽  
Boris R. Belitsky ◽  
Vladimir M. Levdikov ◽  
Anthony J. Wilkinson ◽  
...  
Keyword(s):  
Amino Acids ◽  
Bacillus Subtilis ◽  
Branched Chain Amino Acids ◽  
Site Directed Mutagenesis ◽  
Binding Studies ◽  
Hydrophobic Pocket ◽  
Isoleucine Leucine ◽  
Branched Chain

ABSTRACT Bacillus subtilis CodY protein is a DNA-binding global transcriptional regulator that responds to branched-chain amino acids (isoleucine, leucine, and valine) and GTP. Crystal structure studies have shown that the N-terminal region of the protein includes a GAF domain that contains a hydrophobic pocket within which isoleucine and valine bind. This region is well conserved in CodY homologs. Site-directed mutagenesis was employed to understand the roles of some of the residues in the GAF domain and hydrophobic pocket in interaction with isoleucine and GTP. The F40A, F71E, and F98A forms of CodY were inactive in vivo. They were activatable by GTP but to a much lesser extent by branched-chain amino acids in vitro. The CodY mutant R61A retained partial repression of target promoters in vivo and was able to respond to GTP in vitro but also responded poorly to branched-chain amino acids in vitro unless GTP was simultaneously present. Thus, the GAF domain includes residues essential for full activation of CodY by branched-chain amino acids, but these residues are not critical for activation by GTP. Binding studies with branched-chain amino acids and their analogs revealed that an amino group at position 2 and a methyl group at position 3 of valine are critical components of the recognition of the amino acids by CodY.

scholarly journals Shifts in the Gut Metabolome andClostridium difficileTranscriptome throughout Colonization and Infection in a Mouse Model

mSphere ◽  
2018 ◽  
Vol 3 (2) ◽  
Author(s):  
Joshua R. Fletcher ◽  
Samantha Erwin ◽  
Cristina Lanzas ◽  
Casey M. Theriot
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Clostridium Difficile ◽  
Branched Chain Amino Acids ◽  
Omics Data ◽  
Content Type ◽  
Time Points ◽  
Branched Chain ◽  
Over Time

ABSTRACTAntibiotics alter the gut microbiota and decrease resistance toClostridium difficilecolonization; however, the mechanisms driving colonization resistance are not well understood. Loss of resistance toC. difficilecolonization due to antibiotic treatment is associated with alterations in the gut metabolome, specifically, with increases in levels of nutrients thatC. difficilecan utilize for growthin vitro. To define the nutrients thatC. difficilerequires for colonization and pathogenesisin vivo, we used a combination of mass spectrometry and RNA sequencing (RNA Seq) to model the gut metabolome andC. difficiletranscriptome throughout an acute infection in a mouse model at the following time points: 0, 12, 24, and 30 h. We also performed multivariate-based integration of the omics data to define the signatures that were most important throughout colonization and infection. Here we show that amino acids, in particular, proline and branched-chain amino acids, and carbohydrates decrease in abundance over time in the mouse cecum and thatC. difficilegene expression is consistent with their utilizationin vivo. This was also reinforced by the multivariate-based integration of the omics data where we were able to discriminate the metabolites and transcripts that supportC. difficilephysiology between the different time points throughout colonization and infection. This report illustrates how important the availability of amino acids and other nutrients is for the initial stages ofC. difficilecolonization and progression of disease. Future studies identifying the source of the nutrients and engineering bacteria capable of outcompetingC. difficilein the gut will be important for developing new targeted bacterial therapeutics.IMPORTANCEClostridium difficileis a bacterial pathogen of global significance that is a major cause of antibiotic-associated diarrhea. Antibiotics deplete the indigenous gut microbiota and change the metabolic environment in the gut to one favoringC. difficilegrowth. Here we used metabolomics and transcriptomics to define the gut environment after antibiotics and during the initial stages ofC. difficilecolonization and infection. We show that amino acids, in particular, proline and branched-chain amino acids, and carbohydrates decrease in abundance over time and thatC. difficilegene expression is consistent with their utilization by the bacteriumin vivo. We employed an integrated approach to analyze the metabolome and transcriptome to identify associations between metabolites and transcripts. This highlighted the importance of key nutrients in the early stages of colonization, and the data provide a rationale for the development of therapies based on the use of bacteria that specifically compete for nutrients that are essential forC. difficilecolonization and disease.

scholarly journals Branched‐chain amino acids reduce hepatic iron accumulation and oxidative stress in hepatitis C virus polyprotein‐expressing mice

2014 ◽  
Vol 35 (4) ◽  
pp. 1303-1314 ◽  
Author(s):  
Masaaki Korenaga ◽  
Sohji Nishina ◽  
Keiko Korenaga ◽  
Yasuyuki Tomiyama ◽  
Naoko Yoshioka ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Amino Acids ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Branched Chain Amino Acids ◽  
Iron Accumulation ◽  
Hepatic Iron ◽  
Branched Chain ◽  
Virus Polyprotein ◽  
And Oxidative Stress

Lipidated Methotrexate Microbubbles: A Promising Rheumatoid Arthritis Theranostic Medicine Manipulated via Ultrasonic Irradiation

2021 ◽  
Vol 17 (7) ◽  
pp. 1293-1304
Author(s):  
Zhuofei Zhao ◽  
Xiaona Lin ◽  
Lulu Zhang ◽  
Xia Liu ◽  
Qingwen Wang ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
De Novo ◽  
Low Ph ◽  
Curative Effect ◽  
Inflammatory Microenvironment ◽  
Raw 264.7 Cell ◽  
Infection Focus ◽  
Raw 264.7 Cell Line

De novo designed lipidated methotrexate was synthesized and self-assembled into microbubbles for targeted rheumatoid arthritis theranostic treatment. Controlled lipidatedmethotrexate delivery was achieved by ultrasound-targetedmicrobubble destruction technique. Methotrexate was dissociated inflammatory microenvironment of synovial cavity, owing to representive low pH and enriched leucocyte esterase. We first manipulated methotrexate controlled release with RAW 264.7 cell line in vitro and further verified with rheumatoid arthritis rabbits in vivo. Results showed that lipidated methotrexate microbubbles precisely affected infection focus and significantly enhanced rheumatoid arthritis curative effect comparing with dissociative methotrexate. This study indicates that lipidated methotrexate microbubbles might be considered as a promising rheumatoid arthritis theranostics medicine.

A1151 EFFECT OF BRANCHED CHAIN AMINO ACIDS (BCAA) ON DIAPHRAGM FATIGUE IN VITRO

1990 ◽  
Vol 73 (3A) ◽  
pp. NA-NA
Author(s):  
H. Yamada ◽  
Y. Ohta ◽  
I. Chaudhry ◽  
H. Nagashima ◽  
J. Askanazi ◽  
...  
Keyword(s):  
Amino Acids ◽  
Branched Chain Amino Acids ◽  
Branched Chain ◽  
Diaphragm Fatigue
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