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PloS One 2022To ascertain an appropriate level of isoleucine for LSL-LITE layers (23- to 30-week-old), diets containing total isoleucine concentrations (levels) of 0.66 (Control),...
To ascertain an appropriate level of isoleucine for LSL-LITE layers (23- to 30-week-old), diets containing total isoleucine concentrations (levels) of 0.66 (Control), 0.69, 0.72, 0.75, 0.78, 0.81, and 0.84% were fed as 7 treatments (2730 kcal/kg metabolizable energy) x 7 replicates x 10 birds per replicate. Significance for performance, egg quality, serum biochemistry, and ileal digestibility of protein was determined at P ≤ 0.05. Level, week, and level*week (L*W) were significant for production, egg mass, and feed intake. Level and week were significant for FCR. Week was significant for weight gain. Level was significant for egg weight, specific gravity, and shell thickness; week was also significant for these external egg parameters as well as shape index and proportional shell thickness. L*W was significant for all except shape index. For internal egg measurements, level was significant for proportional yolk, proportional albumen, yolk index, and yolk:albumen. Week was significant for internal egg parameters while L*W significantly affected Haugh unit, proportional albumen weight, yolk index, albumen index, and yolk color. Level was significant for globulin and glucose in serum. Isoleucine at 0.72%, 0.81%, and 0.84% produced the lowest FCR, an important standard in the poultry industry. Considering the low FCR of 1.45 and cost for inclusion as a dietary ingredient, 0.72% isoleucine was chosen for further studies with varying quantities of other branched chain amino acids in diets for young laying hens.
Topics: Isoleucine
PubMed: 35061687
DOI: 10.1371/journal.pone.0261159 -
Current Biology : CB Mar 2020The jasmonate signaling pathway regulates development, growth, and defense responses in plants. Studies in the model eudicot, Arabidopsis thaliana, have identified the...
The jasmonate signaling pathway regulates development, growth, and defense responses in plants. Studies in the model eudicot, Arabidopsis thaliana, have identified the bioactive hormone (jasmonoyl-isoleucine [JA-Ile]) and its Coronatine Insensitive 1 (COI1)/Jasmonate-ZIM Domain (JAZ) co-receptor. In bryophytes, a conserved signaling pathway regulates similar responses but uses a different ligand, the JA-Ile precursor dinor-12-oxo-10,15(Z)-phytodienoic acid (dn-OPDA), to activate a conserved co-receptor. Jasmonate responses independent of JA-Ile and COI1, thought to be mediated by the cyclopentenone OPDA, have also been suggested, but experimental limitations in Arabidopsis have hindered attempts to uncouple OPDA and JA-Ile biosynthesis. Thus, a clear understanding of this pathway remains elusive. Here, we address the role of cyclopentenones in COI1-independent responses using the bryophyte Marchantia polymorpha, which is unable to synthesize JA-Ile but does accumulate OPDA and dn-OPDA. We demonstrate that OPDA and dn-OPDA activate a COI1-independent pathway that regulates plant thermotolerance genes, and consequently, treatment with these oxylipins protects plants against heat stress. Furthermore, we identify that these molecules signal through their electrophilic properties. By performing comparative analyses between M. polymorpha and two evolutionary distant species, A. thaliana and the charophyte alga Klebsormidium nitens, we demonstrate that this pathway is conserved in streptophyte plants and pre-dates the evolutionary appearance of the COI1-dependent jasmonate pathway, which later co-opted the pre-existing dn-OPDA as its ligand. Taken together, our data indicate that cyclopentenone-regulated COI1-independent signaling is an ancient conserved pathway, whose ancestral role was to protect plants against heat stress. This pathway was likely crucial for plants' successful land colonization and will be critical for adaption to current climate warming.
Topics: Arabidopsis; Arabidopsis Proteins; Charophyceae; Cyclopentanes; Gene Expression Regulation, Plant; Genes, Plant; Isoleucine; Marchantia; Oxylipins; Plant Proteins; Signal Transduction; Thermotolerance
PubMed: 32142692
DOI: 10.1016/j.cub.2020.01.023 -
Chemical Research in Toxicology Sep 2016Translesion synthesis (TLS) of the N(2)-2'-deoxyguanosine (dG-N(2)-IQ) adduct of the carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) was investigated in human...
Translesion Synthesis of the N(2)-2'-Deoxyguanosine Adduct of the Dietary Mutagen IQ in Human Cells: Error-Free Replication by DNA Polymerase κ and Mutagenic Bypass by DNA Polymerases η, ζ, and Rev1.
Translesion synthesis (TLS) of the N(2)-2'-deoxyguanosine (dG-N(2)-IQ) adduct of the carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) was investigated in human embryonic kidney 293T cells by replicating plasmid constructs in which the adduct was individually placed at each guanine (G1, G2, or G3) of the NarI sequence (5'-CG1G2CG3CC-3'). TLS efficiency was 38%, 29%, and 25% for the dG-N(2)-IQ located at G1, G2, and G3, respectively, which suggests that dG-N(2)-IQ is bypassed more efficiently by one or more DNA polymerases at G1 than at either G2 or G3. TLS efficiency was decreased 8-35% in cells with knockdown of pol η, pol κ, pol ι, pol ζ, or Rev1. Up to 75% reduction in TLS occurred when pol η, pol ζ, and Rev1 were simultaneously knocked down, suggesting that these three polymerases play important roles in dG-N(2)-IQ bypass. Mutation frequencies (MFs) of dG-N(2)-IQ at G1, G2, and G3 were 23%, 17%, and 11%, respectively, exhibiting a completely reverse trend of the previously reported MF of the C8-dG adduct of IQ (dG-C8-IQ), which is most mutagenic at G3 ( ( 2015 ) Nucleic Acids Res. 43 , 8340 - 8351 ). The major type of mutation induced by dG-N(2)-IQ was targeted G → T, as was reported for dG-C8-IQ. In each site, knockdown of pol κ resulted in an increase in MF, whereas MF was reduced when pol η, pol ι, pol ζ, or Rev1 was knocked down. The reduction in MF was most pronounced when pol η, pol ζ, and Rev1 were simultaneously knocked down and especially when the adduct was located at G3, where MF was reduced by 90%. We conclude that pol κ predominantly performs error-free TLS of the dG-N(2)-IQ adduct, whereas pols η, pol ζ, and Rev1 cooperatively carry out the error-prone TLS. However, in vitro experiments using yeast pol ζ and κ showed that the former was inefficient in full-length primer extension on dG-N(2)-IQ templates, whereas the latter was efficient in both error-free and error-prone extensions. We believe that the observed differences between the in vitro experiments using purified DNA polymerases, and the cellular results may arise from several factors including the crucial roles played by the accessory proteins in TLS.
Topics: DNA Adducts; DNA Replication; DNA-Directed DNA Polymerase; Deoxyguanosine; Diet; HEK293 Cells; Humans; Imidazoles; Isoleucine; Molecular Structure; Mutagens; Nuclear Proteins; Nucleotidyltransferases; Quinoxalines; DNA Polymerase iota
PubMed: 27490094
DOI: 10.1021/acs.chemrestox.6b00221 -
Nutrients Nov 2019The ingestion of whey protein and amino acids with carbohydrate (CHO) enhances the release of glucagon-like peptide-1 (GLP-1) and glucose-dependent-insulinotropic...
BACKGROUND
The ingestion of whey protein and amino acids with carbohydrate (CHO) enhances the release of glucagon-like peptide-1 (GLP-1) and glucose-dependent-insulinotropic peptide (GIP) that promote insulin secretion. It is unknown if L-isoleucine (Ile) and L-leucine (Leu) have this same effect. The purpose of this study was to examine how Ile and Leu influence both GLP-1 and GIP, subsequent pancreatic hormones, and glycemia in healthy, inactive adults.
METHODS
Twelve adults (6F/6M; age 27.4 ± 2 years; BMI 26.3 ± 2 kg/m; lean body mass 53.2 ± 5 kg; body fat 34.1 ± 3%) completed four conditions in a randomized, cross-over fashion. Treatments standardized (0.3 g/kg·LBM) (1) Leu, (2) Ile, (3) Equal (1:1 g) of Leu + Ile, and (4) placebo (Pla, 3.5 g inert stevia) ingested 30 min prior to an oral glucose tolerance test (OGTT). Samples of plasma glucose, insulin, glucagon, GIP, and GLP-1 were assessed.
RESULTS
A treatment ( = 0.01) effect comparing Ile vs. Leu ( = 0.02) in GIP. Area under the curve showed an increase in GIP from Ile compared to Leu and Pla ( = 0.03). No effect was found on GLP-1. The ingestion of Ile prior to CHO augmented GIP concentration greater than Leu or Pla. No correlation was found between GIP, insulin, and glucose between conditions.
CONCLUSIONS
Ile impacts GIP concentration, which did not relate to either insulin or glucose concentrations. Neither Ile, nor Leu seem to have an effect on hyperglycemia ingested prior to a CHO drink.
Topics: Adult; Blood Glucose; Body Composition; Dose-Response Relationship, Drug; Exercise; Female; Humans; Isoleucine; Leucine; Male; Pancreatic Hormones
PubMed: 31689951
DOI: 10.3390/nu11112650 -
Journal of Dairy Science Oct 1976The growth of Streptococcus agalactiae in milk is inhibited by elevated free isoleucine. Free amino acid concentrations in both plasma and milk from three lactating...
The growth of Streptococcus agalactiae in milk is inhibited by elevated free isoleucine. Free amino acid concentrations in both plasma and milk from three lactating dairy cows were studied after intravenous infusions of L-isoleucine at 62, 300, 380, and 840 mumol/kg of body weight. Free isoleucine was lower in milk than in plasma. The concentration of free isoleucine in plasma increased after infusions of isoleucine at the three higher amounts. The highest concentration of free isoleucine in milk measured was 1.5 mumol/100 ml of skin milk. This was observed after infusion of isoleucine for 12 h at 380 mumol/kg of body weight. Free isoleucine in both plasma and milk dropped to the preinfusion concentrations a short time after the isoleucine infusions were stopped. When relatively large quantities of isoleucine were infused into the blood of cows, free isoleucine in blood increased as much as twofold. The source of free isoleucine in milk appeared to be free isoleucine in plasma, but isoleucine in milk only reached concentrations that would give about 50% of the growth inhibition of S. agalactiae obtainable with isoleucine in vitro. It would be difficult to elevate free isoleucine in milk by dietary manipulation sufficiently to inhibit S. agalactiae.
Topics: Amino Acids; Animals; Cattle; Dose-Response Relationship, Drug; Female; Isoleucine; Lactation; Milk; Pregnancy; Streptococcus agalactiae
PubMed: 789418
DOI: 10.3168/jds.S0022-0302(76)84433-5 -
Plant & Cell Physiology Apr 2017Jasmonic acid (JA) is involved in a variety of physiological responses in seed plants. However, the detection and role of JA in lycophytes, a group of seedless vascular...
Jasmonic acid (JA) is involved in a variety of physiological responses in seed plants. However, the detection and role of JA in lycophytes, a group of seedless vascular plants, have remained elusive until recently. This study provides the first evidence of 12-oxo-phytodienoic acid (OPDA), JA and jasmonoyl-isoleucine (JA-Ile) in the model lycophyte Selaginella moellendorffii. Mechanical wounding stimulated the accumulation of OPDA, JA and JA-Ile. These data were corroborated by the detection of enzymatically active allene oxide synthase (AOS), allene oxide cyclase (AOC), 12-oxo-phytodienoic acid reductase 3 (OPR3) and JA-Ile synthase (JAR1) in S. moellendorffii. SmAOS2 is involved in the first committed step of JA biosynthesis. SmAOC1 is a crucial enzyme for generating the basic structure of jasmonates and is actively involved in the formation of OPDA. SmOPR5, a functionally active OPR3-like enzyme, is also vital for the reduction of (+)-cis-OPDA, the only isomer of the JA precursor. The conjugation of JA to Ile by SmJAR1 demonstrates that S. moellendorffii produces JA-Ile. Thus, the four active enzymes have characteristics similar to those in seed plants. Wounding and JA treatment induced the expression of SmAOC1 and SmOPR5. Furthermore, JA inhibited the growth of shoots in S. moellendorffii, which suggests that JA functions as a signaling molecule in S. moellendorffii. This study proposes that JA evolved as a plant hormone for stress adaptation, beginning with the emergence of vascular plants.
Topics: Cyclopentanes; Fatty Acids, Unsaturated; Gene Expression Regulation, Plant; Intramolecular Oxidoreductases; Isoleucine; Ligases; Oxidoreductases; Oxylipins; Plant Proteins; Plant Shoots; Selaginellaceae
PubMed: 28340155
DOI: 10.1093/pcp/pcx031 -
Plant, Cell & Environment Jun 2020Jasmonate synthesis and signalling are essential for plant defense upregulation upon herbivore or microbial attacks. Stress-induced accumulation of jasmonoyl-isoleucine...
Jasmonate synthesis and signalling are essential for plant defense upregulation upon herbivore or microbial attacks. Stress-induced accumulation of jasmonoyl-isoleucine (JA-Ile), the bioactive hormonal form triggering transcriptional changes, is dynamic and transient because of the existence of potent removal mechanisms. Two JA-Ile turnover pathways operate in Arabidopsis, consisting in cytochrome P450 (CYP94)-mediated oxidation and deconjugation by the amidohydrolases IAR3/ILL6. Understanding their impacts was previously blurred by gene redundancy and compensation mechanisms. Here we address the consequences of blocking these pathways on jasmonate homeostasis and defenses in double-2ah, triple-3cyp mutants, and a quintuple-5ko line deficient in all known JA-Ile-degrading activities. These lines reacted differently to either mechanical wounding/insect attack or fungal infection. Both pathways contributed additively to JA-Ile removal upon wounding, but their impairement had opposite impacts on insect larvae feeding. By contrast, only the ah pathway was essential for JA-Ile turnover upon infection by Botrytis, yet only 3cyp was more fungus-resistant. Despite building-up extreme JA-Ile levels, 5ko displayed near-wild-type resistance in both bioassays. Molecular analysis indicated that restrained JA-Ile catabolism resulted in enhanced defense/resistance only when genes encoding negative regulators were not simultaneously overstimulated. This occurred in discrete stress- and pathway-specific combinations, providing a framework for future defense-enhancing strategies.
Topics: Arabidopsis; Botrytis; Cyclopentanes; Feedback, Physiological; Gene Expression Regulation, Plant; Genes, Plant; Genotype; Homeostasis; Isoleucine; Mutation; Oxylipins; RNA, Messenger; Repressor Proteins; Signal Transduction; Stress, Physiological
PubMed: 32162701
DOI: 10.1111/pce.13753 -
Molecules (Basel, Switzerland) Apr 2023Fe (II)-and 2-ketoglutarate-dependent dioxygenases (Fe (II)/α-KG DOs) have been applied to catalyze hydroxylation of amino acids. However, the Fe (II)/α-KG DOs that...
Fe (II)-and 2-ketoglutarate-dependent dioxygenases (Fe (II)/α-KG DOs) have been applied to catalyze hydroxylation of amino acids. However, the Fe (II)/α-KG DOs that have been developed and characterized are not sufficient. L-isoleucine dioxygenase (IDO) is an Fe (II)/α-KG DO that specifically catalyzes the formation of 4-hydroxyisoleucine (4-HIL) from L-isoleucine (L-Ile) and exhibits a substrate specificity toward L-aliphatic amino acids. To expand the substrate spectrum of IDO toward aromatic amino acids, in this study, we analyzed the regularity of the substrate spectrum of IDO using molecular dynamics (MD) simulation and found that the distance between Fe, C2 of α-KG and amino acid chain's C4 may be critical for regulating the substrate specificity of the enzyme. The mutation sites (Y143, S153 and R227) were also subjected to single point saturation mutations based on polarity pockets and residue free energy contributions. It was found that Y143D, Y143I and S153A mutants exhibited catalytic L-phenylalanine activity, while Y143I, S153A, S153Q and S153Y exhibited catalytic L-homophenylalanine activity. Consequently, this study extended the substrate spectrum of IDO with aromatic amino acids and enhanced its application property.
Topics: Amino Acids; Isoleucine; Hydroxylation; Dioxygenases; Phenylalanine; Substrate Specificity
PubMed: 37175159
DOI: 10.3390/molecules28093750 -
International Journal of Biological... Aug 2021Water soluble polymers and their derivatives bound to proteins can dramatically favor the biological activity of new drugs and vaccines. Quantification of the...
Water soluble polymers and their derivatives bound to proteins can dramatically favor the biological activity of new drugs and vaccines. Quantification of the modification degree of the protein is crucial during the development and licensing phase and later in order to monitor the industrial production process and to match product specification. In this work, we describe an innovative way to measure directly the modification degree of polysialylated proteins using proton NMR (Nuclear Magnetic Resonance) spectroscopy. Following a calibration step, the modification degree can be easily deduced by the integration ratio of a separate signal from the polymer and selected signals from the protein. In fact, the upfield-shifted signals of methyl groups from Valine, Leucine and Isoleucine can be used as an internal calibration reference for the integration. In this paper recombinant factor VIII (rFVIII) and recombinant factor IX (rFIX) proteins modified by polysialic acid (PSA) are used to illustrate the accuracy, reproducibility and ease of the method that may replace or complement wet-chemistry approaches.
Topics: Factor IX; Factor VIII; Isoleucine; Leucine; Models, Molecular; Molecular Structure; Protein Conformation; Proton Magnetic Resonance Spectroscopy; Recombinant Proteins; Sialic Acids; Valine
PubMed: 34197856
DOI: 10.1016/j.ijbiomac.2021.06.165 -
Molecules (Basel, Switzerland) Jul 2023As a follow-up to our effort to establish reliable thermodynamic data for amino acids, the heat capacity and phase behavior are reported for -acetyl glycine amide (CAS...
As a follow-up to our effort to establish reliable thermodynamic data for amino acids, the heat capacity and phase behavior are reported for -acetyl glycine amide (CAS RN: 2620-63-5), -acetyl-L-alanine amide (CAS RN: 15962-47-7), -acetyl-L-valine amide (CAS RN: 37933-88-3), -acetyl-L-isoleucine amide (CAS RN: 56711-06-9), and -acetyl-L-leucine amide (CAS RN: 28529-34-2). Prior to heat capacity measurement, thermogravimetric analysis and X-ray powder diffraction were performed to determine decomposition temperatures and initial crystal structures, respectively. The crystal heat capacities of the five -acetyl amino acid amides were measured by Tian-Calvet calorimetry in the temperature interval (266-350 K), by power compensation DSC in the temperature interval (216-471 K), and by relaxation (heat-pulse) calorimetry in the temperature interval (2-268 K). As a result, reference heat capacities and thermodynamic functions for the crystalline phase from 0 K up to 470 K were developed.
Topics: Leucine; Isoleucine; Valine; Amides; Hot Temperature; Amino Acids; Alanine; Glycine
PubMed: 37513312
DOI: 10.3390/molecules28145440