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Scientific Reports Sep 2016Store-operated Ca(2+) entry mediated by STIM1-gated Orai1 channels is essential to activate immune cells and its inhibition or gain-of-function can lead to immune...
Store-operated Ca(2+) entry mediated by STIM1-gated Orai1 channels is essential to activate immune cells and its inhibition or gain-of-function can lead to immune dysfunction and other pathologies. Reactive oxygen species interacting with cysteine residues can alter protein function. Pretreatment of the Ca(2+) selective Orai1 with the oxidant H2O2 reduces ICRAC with C195, distant to the pore, being its major redox sensor. However, the mechanism of inhibition remained elusive. Here we combine experimental and theoretical approaches and show that oxidation of Orai1 leads to reduced subunit interaction, slows diffusion and that either oxidized C195 or its oxidomimetic mutation C195D located at the exit of transmembrane helix 3 virtually eliminates channel activation by intramolecular interaction with S239 of transmembrane helix 4, thereby locking the channel in a closed conformation. Our results demonstrate a novel mechanistic model for ROS-mediated inhibition of Orai1 and identify a candidate residue for pharmaceutical intervention.
Topics: HEK293 Cells; Humans; Mesylates; Mutation; Neoplasm Proteins; ORAI1 Protein; Oxidation-Reduction; Protein Binding; Protein Subunits; Reactive Oxygen Species; Serine; Stromal Interaction Molecule 1; Sulfhydryl Compounds
PubMed: 27624281
DOI: 10.1038/srep33347 -
Plant Disease Nov 2021Fusarium head blight (FHB) primarily caused by is a key disease of small grains. Diseased spikes show symptoms of premature bleaching shortly after infection and have...
Fusarium head blight (FHB) primarily caused by is a key disease of small grains. Diseased spikes show symptoms of premature bleaching shortly after infection and have aborted or shriveled seeds, resulting in reduced yields. The fungus also deteriorates quality and safety of the grain because of production of mycotoxins, especially deoxynivalenol (DON), which can result in grain being docked or rejected at the point of sale. Genetic host resistance to FHB is quantitative, and no complete genetic resistance against this devastating disease is available. Alternative approaches to develop new sources of FHB resistance are needed. In this study, we performed extensive forward genetic screening of the M generation of an ethyl methane sulfonate-induced mutagenized population of cultivar Jagger to isolate variants with FHB resistance. In field testing, 74 mutant lines were found to have resistance against FHB spread, and 30 of these lines also had low DON content. Subsequent testing over 2 years in controlled greenhouse conditions revealed 10 M lines showing significantly lower FHB spread. Seven and 6 of those 10 lines also had reduced DON content and fewer -damaged kernels, respectively. Future endeavors will include identification of the mutations that led to resistance in these variants.
Topics: Ethyl Methanesulfonate; Fusarium; Methane; Plant Diseases; Triticum
PubMed: 34132597
DOI: 10.1094/PDIS-03-21-0670-RE -
Molecular Pharmacology Jun 2021The anesthetic etomidate modulates synaptic 12/32 GABA receptors via binding sites located in transmembrane +/- interfaces. Various approaches indicate that etomidate...
Substituted Cysteine Modification and Protection with -Alkyl- Methanethiosulfonate Reagents Yields a Precise Estimate of the Distance between Etomidate and a Residue in Activated GABA Type A Receptors.
The anesthetic etomidate modulates synaptic 12/32 GABA receptors via binding sites located in transmembrane +/- interfaces. Various approaches indicate that etomidate binds near 2/3M286 side chains, including recent cryogenic electron microscopy images in 122L receptors under nonphysiologic conditions with ∼3.5-Å resolution. We hypothesized that substituted cysteine modification and protection experiments using variably sized -alkyl-methanethiosulfonate (MTS) reagents could precisely estimate the distance between bound etomidate and 3M286 side chains in activated functional receptors. Using voltage-clamp electrophysiology in oocytes expressing 13M286C2L GABA receptors, we measured functional changes after exposing GABA-activated receptors to -alkyl-MTS reagents, from methyl-MTS to -decyl-MTS. Based on previous studies using a large sulfhydryl reagent, we anticipated that cysteine modifications large enough to overlap etomidate sites would cause persistently increased GABA sensitivity and decreased etomidate modulation and that etomidate would hinder these modifications, reducing effects. Based on altered GABA or etomidate sensitivity, ethyl-MTS and larger -alkyl-MTS reagents modified GABA-activated 13M286C2L GABA receptors. Receptor modification by -propyl-MTS or larger reagents caused persistently increased GABA sensitivity and decreased etomidate modulation. Receptor-bound etomidate blocked 3M286C modification by -propyl-MTS, -butyl-MTS, and -hexyl-MTS. In contrast, GABA sensitivity was unaltered by receptor exposure to methyl-MTS or ethyl-MTS, and ethyl-MTS modification uniquely increased etomidate modulation. These results reveal a "cut-on" between ethyl-MTS and -propyl-MTS, from which we infer that --(-propyl) is the smallest 3M286C appendage that overlaps with etomidate sites. Molecular models of the native methionine and --ethyl and --(-propyl) modified cysteines suggest that etomidate is located between 1.7 and 3.0 Å from the 3M286 side chain. SIGNIFICANCE STATEMENT: Precise spatial relationships between drugs and their receptor sites are essential for mechanistic understanding and drug development. This study combined electrophysiology, a cysteine substitution, and -alkyl-methanethiosulfonate modifiers, creating a precise molecular ruler to estimate the distance between a α1β3γ2L GABA type A receptor residue and etomidate bound in the transmembrane +/- interface.
Topics: Anesthetics, Intravenous; Animals; Cysteine; Etomidate; Female; Humans; Indicators and Reagents; Mesylates; Receptors, GABA-A; Xenopus laevis; gamma-Aminobutyric Acid
PubMed: 33766924
DOI: 10.1124/molpharm.120.000224 -
Bioresource Technology Dec 2016The present study aimed to develop thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738 for high lipids production. For this, ethyl methane sulfonate was used,...
The present study aimed to develop thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738 for high lipids production. For this, ethyl methane sulfonate was used, which generated two effective thermo-tolerant mutants, M18 and M24 of Chlorella pyrenoidosa NCIM 2738, capable of surviving at temperature up to 47°C and showing improved lipid and biomass yields. They showed 59.62% and 50.75% increase, respectively in lipid content compared to wild type at 30°C, which could not grow at temperature above 35°C. The novelty of this study lied in incorporation of PAM Flurometry with mutagenesis to generate thermo-tolerant mutants of C. pyrenoidosa and investigating the reasons for increased yields of mutants at cellular and photosynthetic levels with the aim to use them for commercial biodiesel production.
Topics: Biofuels; Biomass; Biotechnology; Chlorella; Ethyl Methanesulfonate; Lipids; Mutagenesis; Photosystem II Protein Complex; Temperature
PubMed: 27689351
DOI: 10.1016/j.biortech.2016.09.049 -
Science (New York, N.Y.) Mar 2019
Topics: Industry; Mesylates; Methane
PubMed: 30898919
DOI: 10.1126/science.aaw7738 -
MSphere Feb 2023Fecal microbiota transplantation (FMT) is an effective means of modulating gut microbiota for the treatment of many diseases, including Clostridioides difficile...
Fecal microbiota transplantation (FMT) is an effective means of modulating gut microbiota for the treatment of many diseases, including Clostridioides difficile infections. The gut-spleen axis has been established, and this is involved in the development and function of the spleen. However, it is not understood whether gut microbiota can be used to improve spleen function, especially in spleens disrupted by a disease or an anti-cancer treatment. In the current investigation, we established that alginate oligosaccharide (AOS)-improved gut microbiota (A10-FMT) can rescue anticancer drug busulfan-disrupted spleen vasculature and spleen function. A10-FMT improved the gene and/or protein expression of genes involved in vasculature development, increased the cell proliferation rate, enhanced the endothelial progenitor cell capability, and elevated the expression of the cell junction molecules to increase the vascularization of the spleen. This investigation found for the first time that the reestablishment of spleen vascularization restored spleen function by improving spleen immune cells and iron metabolism. These findings may be used as a strategy to minimize the side effects of anti-cancer drugs or to improve spleen vasculature-related diseases. Alginate oligosaccharide (AOS)-improved gut microbiota (A10-FMT) can rescue busulfan disrupted spleen vasculature. A10-FMT improved the cell proliferation rate, endothelial progenitor cell capability, and cell junction molecules to increase vasculature formation in the spleen. This reestablishment restored spleen function by improving spleen immune cells and iron metabolism. These findings are useful for the treatment of spleen vasculature-related diseases.
Topics: Busulfan; Spleen; Microbiota; Iron
PubMed: 36511706
DOI: 10.1128/msphere.00581-22 -
Journal of Agricultural and Food... Jun 2022Solubility of growth regulators is essential for their use in agriculture. Four new cytokinin salts─6-benzylaminopurine mesylate (), 6-(2-hydroxybenzylamino)purine...
Solubility of growth regulators is essential for their use in agriculture. Four new cytokinin salts─6-benzylaminopurine mesylate (), 6-(2-hydroxybenzylamino)purine mesylate (), 6-(3-hydroxybenzylamino)purine mesylate (), and 6-(3-methoxybenzylamino)purine mesylate ()─were synthesized, and their crystal structures were determined to clarify structural influence on water solubility. The mesylates were several orders of magnitude more water-soluble than the parent CKs. The new salts significantly reduced chlorophyll degradation and impairment of photosystem II functionality in barley leaf segments undergoing artificial senescence and had pronounced effects on the leaves' endogenous CK pools, maintaining high concentrations of functional metabolites for several days, unlike canonical CKs. A foliar treatment with and increased the harvest yield of spring barley by up to 8% when compared to treatment with the parent CKs while also increasing the number of productive tillers. This effect was attributed to the higher bioavailability of the mesylate salts and the avoidance of dimethyl sulfoxide exposure.
Topics: Cytokinins; Hordeum; Mesylates; Photosynthesis; Plant Leaves; Salts; Water
PubMed: 35658447
DOI: 10.1021/acs.jafc.2c00981 -
ACS Sensors Dec 2023Hyperpolarized (HP) xenon-129 (Xe) magnetic resonance imaging (MRI) has the potential to be used as a molecular imaging modality. For this purpose, numerous...
Hyperpolarized (HP) xenon-129 (Xe) magnetic resonance imaging (MRI) has the potential to be used as a molecular imaging modality. For this purpose, numerous supramolecular cages have been developed and evaluated in the past. Herein, we report a novel and unique macrocycle that can be successfully utilized for xenon MRI, the resorcinarene trimer methanesulfonate (R3-Noria-MeSOH). This molecule is capable of two different contrast mechanisms for xenon-MRI, resulting from an increase in the effective spin-spin relaxation and hyperpolarized chemical exchange saturation transfer (HyperCEST). We have demonstrated a superior negative contrast caused by R3-Noria-MeSOH on HP Xe MRI at 3.0 T as well as HyperCEST imaging of the studied macrocycle. Additionally, we have found that the complex aggregation behaviors of R3-Noria-methanesulfonate and its impact on xenon-129 relaxivity are an area for future study.
Topics: Magnetic Resonance Imaging; Xenon Isotopes; Xenon; Contrast Media; Mesylates
PubMed: 38064687
DOI: 10.1021/acssensors.3c01791 -
International Journal of Radiation... 2021Rice is the prominent crop of world and it frames the important component of food chain. But, long duration of highly preferred rice varieties requires more water and...
PURPOSE
Rice is the prominent crop of world and it frames the important component of food chain. But, long duration of highly preferred rice varieties requires more water and are prone to biotic and abiotic stresses. Short duration rice varieties serve as a promising alternative in such cases. Hence, the present study was taken up to induce early flowering mutants in popular rice variety Bapatla 2231 (BPT 2231), a long duration medium slender rice variety using gamma rays and Ethyl Methane Sulfonate (EMS) treatment combinations.
MATERIALS AND METHODS
Five hundred well filled and viable grains of optimum moisture content (12%) were subjected to irradiation with 8 doses comprising 100 Gy, 200 Gy, 300 Gy, 350 Gy, 400 Gy, 450 Gy, 500 Gy and 600 Gy treatments. For combination treatments, the irradiated seeds of gamma rays were further treated with 30 mM concentrations of Ethyl Methane sulfonate (EMS). The irradiated population were raised up to M generation for the identification of early maturing mutants. The isolated early maturing mutants in M generation were characterized for phenotypic, biochemical and grain qualities.
RESULTS
The frequency of desirable early maturing mutants was observed more in combination treatments than the individual treatments of gamma rays. A total of 34 early maturing mutants (M) with variation for grain quality and biochemical traits were obtained. The mutants M-1, M-3, M-5, M-32, and M-34 were high yielding with fine grain type. The mutants M-4, M-17, M-18, M-19, M-20, and M-26 were high yielding with bold grain type. The mutants M-22, M-23, M-24, M-27, and M-28 were high in nutrient content. The early mutants matured 40-50 days earlier than control.
CONCLUSION
In this variety, the combination treatments showed higher mutation frequency, mutagenic effectiveness and efficiency than the gamma rays in induction of both chlorophyll and viable mutants. This study revealed that the combination treatments were more effective in inducing early mutants than the gamma ray treatments alone. The early maturing mutants can be released as variety after evaluation under different yield trials and the mutants with increased nutrient content and varied grain quality can be utilized in hybridization/heterosis breeding programs for rice crop improvement.
Topics: Ethyl Methanesulfonate; Gamma Rays; Methane; Mutation; Oryza
PubMed: 34597257
DOI: 10.1080/09553002.2021.1987563 -
Cell Proliferation Feb 2024Busulfan is an antineoplastic, which is always accompanied with the abnormal of spermatogonia self-renewal and differentiation. It has been demonstrated that the omega-3...
Busulfan is an antineoplastic, which is always accompanied with the abnormal of spermatogonia self-renewal and differentiation. It has been demonstrated that the omega-3 polyunsaturated fatty acids (PUFAs) benefits mature spermatozoa. However, whether omega-3 can protect endogenous spermatogonia and the detailed mechanisms are still unclear. Evaluate of spermatogenesis function (in vivo) were examined by histopathological analysis, immunofluorescence staining, and western blotting. The levels of lipid metabolites in testicular tissue were determined via liquid chromatography. We investigated the effect of lipid metabolites on Sertoli cells provided paracrine factors to regulate spermatogonia proliferation and differentiation using co-culture system. In our study, we showed that omega-3 PUFAs significantly improved the process of sperm production and elevated the quantity of both undifferentiated Lin28+ spermatogonia and differentiated c-kit+ spermatogonia in a mouse model where spermatogenic function was disrupted by busulfan. Mass spectrometry revealed an increase in the levels of several omega-3 metabolites in the testes of mice fed with omega-3 PUFAs. The eicosapentaenoic acid metabolite 12-hydroxyeicosapentaenoic acid (12-HEPE) up-regulated bone morphogenic protein 4 (BMP4) expression through GPR120-ERK1/2 pathway activation in Sertoli cells and restored spermatogonia proliferation and differentiation. Our study provides evidence that omega-3 PUFAs metabolite 12-HEPE effectively protects spermatogonia and reveals that GPR120 might be a tractable pharmacological target for fertility in men received chemotherapy or severe spermatogenesis dysfunction.
Topics: Humans; Male; Mice; Animals; Busulfan; Semen; Spermatogenesis; Spermatogonia; Spermatozoa; Testis
PubMed: 37743695
DOI: 10.1111/cpr.13551