Exogenous Methionine Research Articles

The Development of LAT1 Efflux Agonists as Mechanistic Probes of Cellular Amino Acid Stress.

Amino acid restriction induces cellular stress and cells often respond via the induction of autophagy. Autophagy or 'self-eating' enables the recycling of proteins and provides the essential amino acids needed for cell survival. Of the naturally occurring amino acids, methionine restriction has pleiotropic effects on cells because methionine also contributes to the intracellular methyl pools required for epigenetic controls as well as polyamine biosynthesis. In this report, we describe the chemical synthesis of four diastereomers of a methionine depletion agent and demonstrate how controlled methionine efflux from cells significantly reduces intracellular methionine, S-adenosylmethionine (SAM), S-adenosyl homocysteine (SAH), and polyamine levels. We also demonstrate that human pancreatic cancer cells respond via a lipid signaling pathway to induce autophagy. The methionine depletion agent causes the large amino acid transporter 1 (LAT1) to preferentially work in reverse and export the cell's methionine (and leucine) stores. The four diastereomers of the lead methionine/leucine depletion agent were synthesized and evaluated for their ability to (a) efflux 3H-leucine from cells, (b) dock to LAT1 in silico, (c) modulate intracellular SAM, SAH, and phosphatidylethanolamine (PE) pools, and (d) induce the formation of the autophagy-associated LC3-II marker. The ability to modulate the intracellular concentration of methionine regardless of exogenous methionine supply provides new molecular tools to better understand cancer response pathways. This information can then be used to design improved therapeutics that target downstream methionine-dependent processes like polyamines.

Comparative study on the utilization of four forms exogenous methionine by red swamp crayfish (Procambarus clarkii)

To investigate the efficacy of four forms of methionine (Met) supplementation in red swamp crayfish (Procambarus clarkii), a Met-deficient all-plant protein diet was used control, along with four supplemental diets: 0.90 g/kg Met-Met, 1.80 g/kg crystal DL-Met, 3.60 g/kg E-Met, and 3.29 g/kg MHA-Ca. The analysis Met + Cys levels of five diets were 10.46 (control), 11.51, 12.50, 12.56, and 10.70 g/kg, respectively. Each diet was hand-fed to five replicates of 18 crayfish each (initial body weight: 4.40 ± 0.02 g) for eight weeks. Supplementation with four forms of exogenous Met resulted in an increase in weight gain rate, specific growth rate, and antioxidant capacity, while decreasing the feed conversion ratio (P < 0.05). The abdominal muscle crude protein content and the protein deposition rate in the Met-Met and E-Met groups were higher than those of the control group (P < 0.05). The deposition rate of Met, arginine, leucine, lysine, threonine, and valine, and the activities of digestive enzymes were higher in the Met-Met group than that of the control group (P < 0.05). The peak times and values of serum biochemical indices were affected by four forms of exogenous Met. Additionally, the hepatopancreatic homocysteine content in the MHA-Ca group was higher than in the control group (P < 0.05). In conclusion, the utilization of Met-Met by Procambarus clarkii was superior to other forms of Met in an all-plant protein diet.

Exogenous methionine contributes to reversing the resistance of Streptococcus suis to macrolides.

Streptococcus suis (S. suis) has been increasingly recognized as a porcine zoonotic pathogen that threatens the health of both pigs and humans. Multidrug-resistant Streptococcus suis is becoming increasingly prevalent, and novel strategies to treat bacterial infections caused by these organisms are desperately needed. In the present study, an untargeted metabolomics analysis showed that the significant decrease in methionine content and the methionine biosynthetic pathway were significantly affected by the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis in drug-resistant S. suis. The addition of L-methionine restored the bactericidal activity of macrolides, doxycycline, and ciprofloxacin on S. suis in vivo and in vitro. Further studies showed that the exogenous addition of methionine affects methionine metabolism by reducing S-adenosylmethionine synthetase activity and the contents of S-adenosylmethionine, S-adenosyl homocysteine, and S-ribose homocysteine. Methionine can decrease the total methylation level and methylesterase activity in multidrug resistant S. suis. The drug transport proteins and efflux pump genes were significantly downregulated in S. suis by exogenous L-methionine. Moreover, the exogenous addition of methionine can reduce the survival of S. suis by affecting oxidative stress and metal starvation in bacteria. Thus, L-methionine may influence the development of resistance in S. suis through methyl metabolism and metal starvation. This study provides a new perspective on the mitigation of drug resistance in S. suis.IMPORTANCEBacterial antibiotic resistance has become a severe threat to human and animal health. Increasing the efficacy of existing antibiotics is a promising strategy against antibiotic resistance. Here, we report that L-methionine enhances the efficacy of macrolides, doxycycline, and ciprofloxacin antibiotics in killing Streptococcus suis, including multidrug-resistant pathogens. We investigated the mechanism of action of exogenous methionine supplementation in restoring macrolides in Streptococcus suis and the role of the methionine cycle pathway on methylation levels, efflux pump genes, oxidative stress, and metal starvation in Streptococcus suis. It provides a theoretical basis for the rational use of macrolides in clinical practice and also identifies a possible target for restoring drug resistance in Streptococcus suis.

Asymmetric and symmetric protein arginine methylation in methionine-addicted human cancer cells.

The methionine addiction of cancer cells is known as the Hoffman effect. While non-cancer cells in culture can utilize homocysteine in place of methionine for cellular growth, most cancer cells require exogenous methionine for proliferation. It has been suggested that a biochemical basis of this effect is the increased utilization of methionine for S-adenosylmethionine, the major methyl donor for a variety of cellular methyltransferases. Recent studies have pointed to the role of S-adenosylmethionine-dependent protein arginine methyltransferases (PRMTs) in cell proliferation and cancer. To further understand the biochemical basis of the methionine addiction of cancer cells, we compared protein arginine methylation in two previously described isogenic cell lines, a methionine-addicted 143B human osteosarcoma cell line and its less methionine-dependent revertant. Previous work showed that the revertant cells were significantly less malignant than the parental cells. In the present study, we utilized antibodies to detect the asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) products of PRMTs in polypeptides from cellular extracts and purified histone preparations of these cell lines fractionated by SDS-PAGE. Importantly, we observed little to no differences in the banding patterns of ADMA- and SDMA-containing species between the osteosarcoma parental and revertant cell lines. Furthermore, enzymatic activity assays using S-adenosyl-ʟ-[methyl-3H] methionine, recombinantly purified PRMT enzymes, cell lysates, and specific PRMT inhibitors revealed no major differences in radiolabeled polypeptides on SDS-PAGE gels. Taken together, these results suggest that changes in protein arginine methylation may not be major contributors to the Hoffman effect and that other consequences of methionine addiction may be more important in the metastasis and malignancy of osteosarcoma and potentially other cancers.

Application of Methionine Increases the Germination Rate of Maize Seeds by Triggering Multiple Phenylpropanoid Biosynthetic Genes at Transcript Levels.

Methionine is an essential amino acid that initiates protein synthesis and serves as a substrate for various chemical reactions. Methionine metabolism plays an important role in Arabidopsis seed germination, but how methionine works in seed germination of maize has not been elucidated. We compared the changes in germination rate, the contents of methionine and folates, and transcriptional levels using transcriptome analysis under water or exogenous methionine treatment. The results indicate that the application of methionine increases seed germination rate (95% versus 70%), leading to significant differences in the content of methionine at 36 h, which brought the rapid increase forward by 12 h in the embryo and endosperm. Transcriptome analysis shows that methionine mainly affects the proliferation and differentiation of cells in the embryo, and the degradation of storage substances and signal transduction in the endosperm. In particular, multiple phenylpropanoid biosynthetic genes were triggered upon methionine treatment during germination. These results provide a theoretical foundation for promoting maize seed germination and serve as a valuable theoretical resource for seed priming strategies.

Genomic footprinting uncovers global transcription factor responses to amino acids in Escherichia coli.

Our knowledge of transcriptional responses to changes in nutrient availability comes primarily from few well-studied transcription factors (TFs), often lacking an unbiased genome-wide perspective. Leveraging recent advances allowing bacterial genomic footprinting, we comprehensively mapped the genome-wide regulatory responses of Escherichia coli to exogenous leucine, methionine, alanine, and lysine. The global TF Lrp was found to individually sense three amino acids and mount three different target gene responses. Overall, 531 genes had altered RNA polymerase occupancy, and 32 TFs responded directly or indirectly to the presence of amino acids, including regulators of membrane and osmotic pressure homeostasis. About 70% of the detected TF-DNA interactions had not been reported before. We thus identified 682 previously unknown TF-binding locations, for a subset of which the involved TFs were identified by affinity purification. This comprehensive map of amino acid regulation illustrates the incompleteness of the known transcriptional regulation network, even in E.coli.

Cytotoxic Effect of L-Methioninase from Brevibacterium linens BL2 in Combination with Etoposide against Glioblastoma Cells

L-methioninase degrades methionine, which is essential in methionine-dependent cancer cells, resulting in specific cell death. Normal cells can synthesize their own methionine amino acids even in the absence of exogenous methionine. This selective targeting of cancer cells makes L-methioninase a promising therapeutic candidate for cancer. In this study, L-methioninase was partially purified from Brevibacterium linens BL2. The specific activity of the enzyme was found as 3.055 units/mg. IC50 values (24 h) of the enzyme were 5.792 units/mL for U87MG cell line and 5.215 units/mL for T98G cell line. When L-methioninase and etoposide were used in combination, synergistic cytotoxic and cell migration inhibition effects on U87MG and T98G cells alongside decreased cytotoxic activity on the Mouse Embryonic Fibroblast and HaCaT cells compared to etoposide alone were observed. Additionally, colony numbers of U87MG cells were significantly reduced by L-methioninase and etoposide administration after 21 days of incubation. Furthermore, L-methioninase suppressed the expression levels of survivin and c-Myc while increasing the expression level of Caspase-3 in both glioblastoma cell lines. These effects were enhanced when etoposide was used in combination with etoposide. This investigation reveals that the L-methioninase enzyme not only exhibited cytotoxic effects on U87MG and T98G cells but also enhanced the anti-proliferative effects of etoposide when used in combination while also demonstrating fewer adverse effects on normal cells.

Biostimulants-Based Amino Acids Augment Physio-Biochemical Responses and Promote Salinity Tolerance of Lettuce Plants (Lactuca sativa L.)

Studying the biostimulation effect of amino acids indicated their possible role in salt stress mitigation. In this investigation, six exogenous amino acids (alanine (Ala), arginine (Arg), glutamine (Glu), glycine (Gly), methionine (Met), and proline (Pro)) at 0.5 g/L were sprayed to evaluate their impact on lettuce plants cultivated under simulated salt stress conditions. Photosynthetic pigments, ion absorption, endogenous amino acids contents, catalase (CAT), and peroxidase (POD) enzyme activities were determined. A significant alleviation of salt stress was noticed when EAAs were used in the stress-induced plants, and applying Gly, Met, and Pro improved the plant status under salt stress conditions. The highest electric conductivity (568 μS/g) was testified from the control treatment (50 mM NaCl), while applying exogenous amino acids reduced electrical conductivity (EC), and the result was located between 469 and 558 μS/g. AAs alleviated Cl- anions in the lettuce leaves by 25% in comparison to control plants. Na+ cations were alleviated when the stress-induced plants were sprayed with amino acids. In contrast, applying amino acids promoted K+ uptake, and Arg presented the highest contents (3226 μg/g). AAs promoted chlorophyll (chl a and chl b) concentrations compared to the control treatment, and Met produced the maximum chl a content, while the carotene (car) contents significantly augmented when Gly, Met, and Pro were applied. AAs were highly generated in non-stressed treatment (Std) compared to the control. Under simulated salinity stress, Met and Pro application enhanced proteinogenic amino acids expression. Compared to Ctl treatment, peroxidase enzyme activities significantly diminished in the other treatments, which fell by over 40% when Gly, Met, and Pro were sprayed.

Radical-Driven Methane Formation in Humans Evidenced by Exogenous Isotope-Labeled DMSO and Methionine.

Methane (CH4), which is produced endogenously in animals and plants, was recently suggested to play a role in cellular physiology, potentially influencing the signaling pathways and regulatory mechanisms involved in nitrosative and oxidative stress responses. In addition, it was proposed that the supplementation of CH4 to organisms may be beneficial for the treatment of several diseases, including ischemia, reperfusion injury, and inflammation. However, it is still unclear whether and how CH4 is produced in mammalian cells without the help of microorganisms, and how CH4 might be involved in physiological processes in humans. In this study, we produced the first evidence of the principle that CH4 is formed non-microbially in the human body by applying isotopically labeled methylated sulfur compounds, such as dimethyl sulfoxide (DMSO) and methionine, as carbon precursors to confirm cellular CH4 formation. A volunteer applied isotopically labeled (2H and 13C) DMSO on the skin, orally, and to blood samples. The monitoring of stable isotope values of CH4 convincingly showed the conversion of the methyl groups, as isotopically labeled CH4 was formed during all experiments. Based on these results, we considered several hypotheses about endogenously formed CH4 in humans, including physiological aspects and stress responses involving reactive oxygen species (ROS). While further and broader validation studies are needed, the results may unambiguously serve as a proof of concept for the endogenous formation of CH4 in humans via a radical-driven process. Furthermore, these results might encourage follow-up studies to decipher the potential physiological role of CH4 and its bioactivity in humans in more detail. Of particular importance is the potential to monitor CH4 as an oxidative stress biomarker if the observed large variability of CH4 in breath air is an indicator of physiological stress responses and immune reactions. Finally, the potential role of DMSO as a radical scavenger to counteract oxidative stress caused by ROS might be considered in the health sciences. DMSO has already been investigated for many years, but its potential positive role in medical use remains highly uncertain.

Selective Methionine Pool Exhaustion Mediated by a Sequential Positioned MOF Nanotransformer for Intense Cancer Immunotherapy.

Cancer cells are addictive to exogenous methionine to gear toward tumor proliferation. Meanwhile, they could replenish methionine pool from polyamine metabolism through methionine salvage pathway. However, the current developed therapeutic tactics for methionine depletion remain facing great challenges in terms of the selectivity, safety and efficiency. Herein, a sequential positioned metal organic framework (MOF) nanotransformer was designed to selectively exhaust methionine pool via inhibiting the uptake of methionine and throttling its salvage pathway for enhanced cancer immunotherapy. The MOF nanotransformer could restrain the open source and reduce the reflux of methionine to exhaust methionine pool of cancer cells. Moreover, the intracellular traffic routes of sequential positioned MOF nanotransformer matched well with the distribution of polyamines, which was conducive to the oxidation of polyamines via its responsive deformability and nanozyme-augmented Fenton-like reaction for the final exhaustion of intracellular methionine. Our results verified that the well-designed platform could not only kill cancer cells efficiently but also promote the infiltration of CD8 and CD4 T cells for intensive cancer immunotherapy. Overall, we believe that our work will inspire the construction of novel MOF-based antineoplastic platform and provide new insights into the development of metabolic-related immunotherapy. This article is protected by copyright. All rights reserved.

Synergy of Combining Methionine Restriction and Chemotherapy: The Disruptive Next Generation of Cancer Treatment.

All cancer cell types are methionine-addicted, which is termed the Hoffman effect. Cancer cells, unlike normal cells, cannot survive without large amount of methionine. In general, when methionine is depleted, both normal cells and cancer cells synthesize methionine from homocysteine, but cancer cells consume large amounts of methionine and they cannot survive without exogenous methionine. For this reason, methionine restriction has been shown to be effective against many cancers in vitro and in vivo. Methionine restriction arrests cancer cells in the S/G2-phase of the cell cycle. Cytotoxic agents that act in the S/G2-phase are highly effective when used in combination with methionine restriction due to the cancer cells being trapped in S/G2-phase, unlike normal cells which arrest in G1/G0-phase. Combining methionine restriction and chemotherapeutic drugs for cancer treatment is termed the Hoffman protocol. The efficacy of many cytotoxic agents and molecular-targeted drugs in combination with methionine restriction has been demonstrated. The most effective method of methionine restriction is the administration of recombinant methioninase (rMETase), which degrades methionine. The efficacy of rMETase has been reported in mice and human patients by oral administration. The present review describes studies on anticancer drugs that showed synergistic efficacy in combination with methionine restriction, including rMETase administration. It is proposed that the next disruptive generation of cancer chemotherapy should employ current therapy in combination with methionine restriction for all cancer types.

Abstract 6045: Role of PP2A methylation on methionine dependence of cancer

Abstract Dependence on exogenous methionine is a well-known, but poorly understood cancer-specific metabolic requirement. Although most cancer cells can convert homocysteine to methionine, they are unable to grow when methionine depleted media is supplemented with homocysteine. In contrast, non-tumorigenic cells proliferate well in homocysteine media. Previous work from our laboratory has shown that cancer cells cultured in homocysteine medium increase metabolic flux towards the transsulfuration pathway, away from S-adenosylmethionine (SAM) synthesis resulting in overall reduced methylation potential. We have shown that methionine sensing occurs independently of the mTOR pathway, which is known to link amino acid abundance and cell growth. Instead, our results suggest that Protein Phosphatase 2A (PP2A) methylation is sensitive to changes in the SAM/SAH ratio. Reduced PP2A methylation correlates with reduced cell proliferation in homocysteine medium. Furthermore, overexpression of the PP2A demethylase Pme1 forcibly reduces PP2A methylation and induces methionine dependence in cells that normally proliferate in homocysteine medium. These findings suggest a model that links methionine metabolism to cell cycle progression via regulation of PP2A methylation. Here we use quantitative proteomics to define changes in B-subunit binding to PP2A during methionine starvation to investigate the molecular mechanism of this methionine dependence seen in cancer. Citation Format: Anna Andronicos, Da-Wei Lin, Yi-Chen Su, Mari Ishak Gabra, Mei Kong, Peter Kaiser. Role of PP2A methylation on methionine dependence of cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6045.

Disruption of the metC Gene Affects Methionine Biosynthesis in Pectobacterium carotovorum subsp. carotovorum Pcc21 and Reduces Soft-Rot Disease.

Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.

Methionine Restriction Increases Exosome Production and Secretion in Breast Cancer Cells.

Methionine addiction is the elevated requirement for exogenous methionine for growth and survival of cancer cells, termed the Hoffman effect. Methionine-addicted cancer cells synthesize normal or excess amounts of methionine but still need an external source of methionine. Methionine restriction (MR) by either a methionine-free medium or in vivo by a low-methionine diet or by methioninase, selectively arrests cancer cells in the late S/G2 cell cycle phase, but not normal cells. The present study focuses on the comparison of production and secretion of exosomes by cancer cells under MR and normal conditions. MDA-MB-231 cells (triple-negative breast cancer), containing exosomes labeled with CD63-GFP (CD63-GFP exosomes), were visualized by fluorescence microscopy. MDA-MB-231 cells expressing exosome-specific CD63-GFP were cultured in methionine-containing (MET+) or in methionine-free (MET-) DMEM conditions. Exosomes were isolated from conditioned medium of cultured MDA-MD-231 cells by ultracentrifugation and characterized by nanoparticle tracking analysis (NTA) and Western blotting. When MDA-MB-231-CD63-GFP cells were cultured under MR conditions, they arrested their growth and CD63-GFP-expressing exosomes were strongly increased in the cells. MR resulted in approximately a 2-fold increase in exosome production and secretion per cell, even though cell growth was arrested. Methionine restriction thus resulted in elevated exosome production and secretion per surviving cell. Exosome production and secretion in the cancer cells increased under MR, suggesting a relation between MR and exosome production and secretion.

On the illusion of auxotrophy: met15Δ yeast cells can grow on inorganic sulfur, thanks to the previously uncharacterized homocysteine synthase Yll058w

Organisms must either synthesize or assimilate essential organic compounds to survive. The homocysteine synthase Met15 has been considered essential for inorganic sulfur assimilation in yeast since its discovery in the 1970s. As a result, MET15 has served as a genetic marker for hundreds of experiments that play a foundational role in eukaryote genetics and systems biology. Nevertheless, we demonstrate here through structural and evolutionary modeling, invitro kinetic assays, and genetic complementation, that an alternative homocysteine synthase encoded by the previously uncharacterized gene YLL058W enables cells lacking Met15 to assimilate enough inorganic sulfur for survival and proliferation. These cells however fail to grow in patches or liquid cultures unless provided with exogenous methionine or other organosulfurs. We show that this growth failure, which has historically justified the status of MET15 as a classic auxotrophic marker, is largely explained by toxic accumulation of the gas hydrogen sulfide because of a metabolic bottleneck. When patched or cultured with a hydrogen sulfide chelator, and when propagated as colony grids, cells without Met15 assimilate inorganic sulfur and grow, and cells with Met15 achieve even higher yields. Thus, Met15 is not essential for inorganic sulfur assimilation in yeast. Instead, MET15 is the first example of a yeast gene whose loss conditionally prevents growth in a manner that depends on local gas exchange. Our results have broad implications for investigations of sulfur metabolism, including studies of stress response, methionine restriction, and aging. More generally, our findings illustrate how unappreciated experimental variables can obfuscate biological discovery.

Evaluation of dietary selenium methionine levels and their effects on growth performance, antioxidant status, and meat quality of intensively reared juvenile Hypophthalmichthys molitrix.

The objective of this study was to optimize the organic selenium (Se) requirements of intensively reared silver carp (Hypophthalmichthys molitrix). A total of n = 300 juveniles silver carp 11.40±0.52 cm long, and average weighing 25.28±0.18 grams were randomly assigned to 15 aquaria (20 fish/100L aquaria) and subjected to five different dietary Se levels in a completely randomized design. The diets were pelleted supplemented with exogenous Se methionine @ 0.0, 0.3, 0.6, 0.9 and 1.2 mg/kg of the diet. The fourteen days of aquaria acclimatization was given to fish and then an 84-day feeding trial was conducted. The group supplemented with 0.9 mg/kg Se had greater feed intake, gain in length, body weight %, and specific growth rate with a better feed conversion ratio as compared to those fed on the rest of the dietary levels or control (P<0.05). The deposition of Se was greater in the liver, and kidneys of the fishes fed on diets containing 0.9 and 1.2 mg Se levels than in the rest of the treatments (P<0.05). However, dietary Se levels had no effects on the bioaccumulation of Se in muscle tissues (P>0.05). The proximate analysis showed that dry matter, crude protein, and fat contents of meat were not changed (P>0.05) by dietary treatments. Similarly, values of TBARS, RBCs, Hb, and blood glucose contents were similar (P>0.05) across the treatments. However, the concentration of WBCs, HCT, and MCHC was greater in those groups fed on 0.9 and 1.2 Se levels than in those fed on 0.6, 0.3, and 0.0 Se levels respectively (P<0.05). The activities of ALT, AST, and ALP were lowered in the 0.9 mg Se supplemented fishes compared with the rest of the treatments (P<0.05). The SOD, catalases, and GPx levels for muscle, liver, and whole body were greater (P<0.05) in the Se-supplemented groups than in the control. These outcomes indicated that up to 0.9 mg/kg inclusion of methionine-based Se in the diet of juvenile silver carp improved the growth performance, feed conversion ratio, organs Se enrichment, and antioxidant status without any compromise on meat quality.

Exogenous Proline, Methionine, and Melatonin Stimulate Growth, Quality, and Drought Tolerance in Cauliflower Plants

The impact of proline, methionine, and melatonin on cauliflower plants under drought stress is still unclear in the available publications. So, this research aims to study these biochemical compounds’ effects on cauliflower plants grown under well-irrigated and drought-stressed conditions. The obtained results showed that under drought-stressed conditions, foliar application of proline, methionine, and melatonin significantly (p ≤ 0.05) enhanced leaf area, leaf chlorophyll content, leaf relative water content (RWC), vitamin C, proline, total soluble sugar, reducing sugar, and non-reducing sugar compared to the untreated plants. These treatments also significantly increased curd height, curd diameter, curd freshness, and dry matter compared to untreated plants. Conversely, the phenolic-related enzymes including polyphenol oxidase (PPO), peroxidase (POD), and phenylalanine ammonia-lyase (PAL) were significantly reduced compared to the untreated plants. A similar trend was observed in glucosinolates, abscisic acid (ABA), malondialdehyde (MDA), and total phenols. Eventually, it can be concluded that the foliar application of proline, methionine, and melatonin can be considered a proper strategy for enhancing the growth performance and productivity of cauliflower grown under drought-stressed conditions.

Mechanisms of chlorate toxicity and resistance in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic bacterial pathogen that often encounters hypoxic/anoxic environments within the host, which increases its tolerance to many conventional antibiotics. Toward identifying novel treatments, we explored the therapeutic potential of chlorate, a pro-drug that kills hypoxic/anoxic, antibiotic-tolerant P. aeruginosa populations. While chlorate itself is relatively nontoxic, it is enzymatically reduced to the toxic oxidizing agent, chlorite, by hypoxically induced nitrate reductase. To better assess chlorate's therapeutic potential, we investigated mechanisms of chlorate toxicity and resistance in P. aeruginosa. We used transposon mutagenesis to identify genes that alter P. aeruginosa fitness during chlorate treatment, finding that methionine sulfoxide reductases (Msr), which repair oxidized methionine residues, support survival during chlorate stress. Chlorate treatment leads to proteome-wide methionine oxidation, which is exacerbated in a ∆msrA∆msrB strain. In response to chlorate, P. aeruginosa upregulates proteins involved in a wide range of functions, including metabolism, DNA replication/repair, protein repair, transcription, and translation, and these newly synthesized proteins are particularly vulnerable to methionine oxidation. The addition of exogenous methionine partially rescues P. aeruginosa survival during chlorate treatment, suggesting that widespread methionine oxidation contributes to death. Finally, we found that mutations that decrease nitrate reductase activity are a common mechanism of chlorate resistance.

How Different Dietary Methionine Sources Could Modulate the Hepatic Metabolism in Rainbow Trout?

In aquafeeds in which plant proteins are used to replace fishmeal, exogenous methionine (Met) sources are demanded to balance the amino acid composition of diets and meet the metabolic fish requirements. Nonetheless, since different synthetic Met sources are commercially available, it is important to determine their bioavailability and efficacy. To address this issue, we conducted a two-month feeding trial with rainbow trout (Oncorhynchus mykiss), which were fed diets supplemented with five different forms of Met: Met-Met, L-Met, HMTBa, DL-Met, and Co DL-Met. No differences in growth performance were found in trout fed with different Met forms, but changes in the whole-body composition were found. In particular, Met-Met and L-Met promoted a significant body lipid reduction, whereas the protein retention was significantly increased in fish fed with HMTBa and Co DL-Met. The latter affected the hepatic Met metabolism promoting the trans-sulfuration pathway through the upregulation of CBS gene expression. Similarly, the L-Met enhanced the remethylation pathway through an increase in BHMT gene expression to maintain the cellular demand for Met. Altogether, our findings suggest an optimal dietary intake of all tested Met sources with similar promoting effects on fish growth and hepatic Met metabolism. Nevertheless, the mechanisms underlying these effects warrant further investigation.

Glioblastoma and Methionine Addiction.

Glioblastoma is a fatal brain tumor with a bleak prognosis. The use of chemotherapy, primarily the alkylating agent temozolomide, coupled with radiation and surgical resection, has provided some benefit. Despite this multipronged approach, average patient survival rarely extends beyond 18 months. Challenges to glioblastoma treatment include the identification of functional pharmacologic targets as well as identifying drugs that can cross the blood-brain barrier. To address these challenges, current research efforts are examining metabolic differences between normal and tumor cells that could be targeted. Among the metabolic differences examined to date, the apparent addiction to exogenous methionine by glioblastoma tumors is a critical factor that is not well understood and may serve as an effective therapeutic target. Others have proposed this property could be exploited by methionine dietary restriction or other approaches to reduce methionine availability. However, methionine links the tumor microenvironment with cell metabolism, epigenetic regulation, and even mitosis. Therefore methionine depletion could result in complex and potentially undesirable responses, such as aneuploidy and the aberrant expression of genes that drive tumor progression. If methionine manipulation is to be a therapeutic strategy for glioblastoma patients, it is essential that we enhance our understanding of the role of methionine in the tumor microenvironment.