Metabolic rewiring is a characteristic hallmark of cancer cells. This phenomenon sustains uncontrolled proliferation and resistance to apoptosis by increasing nutrients and energy supply. However, reprogramming comes together with vulnerabilities that can be used against tumor and can be applied in targeted therapy. In the last years, the genetic background of tumors has been identified thoroughly and new therapies targeting those mutations tested. Nevertheless, we propose that targeting the phenotype of cancer cells could be another way of treatment aiming to avoid drug resistance and non-responsiveness of cancer patients. Amino acid metabolism is part of the altered processes in cancer cells. Amino acids are building blocks and also sensors of signaling pathways regulating main biological processes. In this comprehensive review, we described four amino acids (asparagine, arginine, methionine, and cysteine) which have been actively investigated as potential targets for anti-tumor therapy. Asparagine depletion is successfully used for decades in the treatment of acute lymphoblastic leukemia and there is a strong implication to apply it to other types of tumors. Arginine auxotrophic tumors are great candidates for arginine-starvation therapy. Higher requirement for essential amino acids such as methionine and cysteine point out promising targetable weaknesses of cancer cells.
- MeSH
- aminokyseliny metabolismus MeSH
- arginin metabolismus terapeutické užití MeSH
- asparagin * metabolismus terapeutické užití MeSH
- cystein metabolismus MeSH
- lidé MeSH
- methionin MeSH
- nádory * metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Extravasation of monocytes into tissue and to the site of injury is a fundamental immunological process, which requires rapid responses via post translational modifications (PTM) of proteins. Protein arginine methyltransferase 7 (PRMT7) is an epigenetic factor that has the capacity to mono-methylate histones on arginine residues. Here we show that in chronic obstructive pulmonary disease (COPD) patients, PRMT7 expression is elevated in the lung tissue and localized to the macrophages. In mouse models of COPD, lung fibrosis and skin injury, reduced expression of PRMT7 associates with decreased recruitment of monocytes to the site of injury and hence less severe symptoms. Mechanistically, activation of NF-κB/RelA in monocytes induces PRMT7 transcription and consequential mono-methylation of histones at the regulatory elements of RAP1A, which leads to increased transcription of this gene that is responsible for adhesion and migration of monocytes. Persistent monocyte-derived macrophage accumulation leads to ALOX5 over-expression and accumulation of its metabolite LTB4, which triggers expression of ACSL4 a ferroptosis promoting gene in lung epithelial cells. Conclusively, inhibition of arginine mono-methylation might offer targeted intervention in monocyte-driven inflammatory conditions that lead to extensive tissue damage if left untreated.
- MeSH
- arginin metabolismus MeSH
- chronická obstrukční plicní nemoc * genetika MeSH
- histony metabolismus MeSH
- intracelulární signální peptidy a proteiny MeSH
- lidé MeSH
- monocyty metabolismus MeSH
- myši MeSH
- proteinarginin-N-methyltransferasy * metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
We adapted a radioligand receptor binding assay for measuring insulin levels in unknown samples. The assay enables rapid and accurate determination of insulin concentrations in experimental samples, such as from insulin-secreting cells. The principle of the method is based on the binding competition of insulin in a measured sample with a radiolabeled insulin for insulin receptor (IR) in IM-9 cells. Both key components, radiolabeled insulin and IM-9 cells, are commercially available. The IR binding assay was used to determine unknown amounts of insulin secreted by MIN6 β cell line after stimulation with glucose, arginine, ornithine, dopamine, and serotonin. The experimental data obtained by the IR binding assay were compared to the results determined by RIA kits and both methods showed a very good agreement of results. We observed the stimulation of glucose-induced insulin secretion from MIN6 cells by arginine, weaker stimulation by ornithine, but inhibitory effects of dopamine. Serotonin effects were either stimulatory or inhibitory, depending on the concentration of serotonin used. The results will require further investigation. The study also clearly revealed advantages of the IR binding assay that allows the measuring of a higher throughput of measured samples, with a broader range of concentrations than in the case of RIA kits. The IR binding assay can provide an alternative to standard RIA and ELISA assays for the determination of insulin levels in experimental samples and can be especially useful in scientific laboratories studying insulin production and secretion by β cells and searching for new modulators of insulin secretion.
- MeSH
- arginin metabolismus MeSH
- beta-buňky metabolismus MeSH
- buněčné linie MeSH
- dopamin metabolismus MeSH
- glukosa metabolismus MeSH
- inzulin analýza metabolismus MeSH
- krysa rodu rattus MeSH
- Langerhansovy ostrůvky metabolismus MeSH
- lidé MeSH
- myši MeSH
- ornithin metabolismus MeSH
- potkani Wistar MeSH
- radioimunoanalýza metody MeSH
- radioligandová zkouška metody MeSH
- sekrece inzulinu * MeSH
- serotonin metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Hexameric arginine repressor, ArgR, is the feedback regulator of bacterial L-arginine regulons, and sensor of L-arg that controls transcription of genes for its synthesis and catabolism. Although ArgR function, as well as its secondary, tertiary, and quaternary structures, is essentially the same in E. coli and B. subtilis, the two proteins differ significantly in sequence, including residues implicated in the response to L-arg. Molecular dynamics simulations are used here to evaluate the behavior of intact B. subtilis ArgR with and without L-arg, and are compared with prior MD results for a domain fragment of E. coli ArgR. Relative to its crystal structure, B. subtilis ArgR in absence of L-arg undergoes a large-scale rotational shift of its trimeric subassemblies that is very similar to that observed in the E. coli protein, but the residues driving rotation have distinct secondary and tertiary structural locations, and a key residue that drives rotation in E. coli is missing in B. subtilis. The similarity of trimer rotation despite different driving residues suggests that a rotational shift between trimers is integral to ArgR function. This conclusion is supported by phylogenetic analysis of distant ArgR homologs reported here that indicates at least three major groups characterized by distinct sequence motifs but predicted to undergo a common rotational transition. The dynamic consequences of L-arg binding for transcriptional activation of intact ArgR are evaluated here for the first time in two-microsecond simulations of B. subtilis ArgR. L-arg binding to intact B. subtilis ArgR causes a significant further shift in the angle of rotation between trimers that causes the N-terminal DNA-binding domains lose their interactions with the C-terminal domains, and is likely the first step toward adopting DNA-binding-competent conformations. The results aid interpretation of crystal structures of ArgR and ArgR-DNA complexes.
- MeSH
- alosterická regulace MeSH
- arginin chemie metabolismus MeSH
- Bacillus subtilis chemie genetika metabolismus MeSH
- bakteriální proteiny chemie genetika metabolismus MeSH
- entropie MeSH
- Escherichia coli chemie genetika metabolismus MeSH
- fylogeneze MeSH
- konformace proteinů, alfa-helix MeSH
- konformace proteinů, beta-řetězec MeSH
- proteinové domény MeSH
- regulon genetika MeSH
- represorové proteiny chemie genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- simulace molekulární dynamiky MeSH
- vazba proteinů MeSH
- vodíková vazba MeSH
- Publikační typ
- časopisecké články MeSH
High-salt intake is one of the major dietary determinants of increased blood pressure and cardiovascular disease. Thus, there is scientific and medical interest in understanding the mechanistic abnormalities mediating the pressor effects of salt (salt sensitivity). According to historical theory, salt sensitivity stems from an impairment in renal function (referred to as "abnormal pressure natriuresis" or a "natriuretic handicap"), which causes salt-sensitive subjects to excrete a sodium load more slowly, and retain more of it than salt-resistant normotensive controls. However, this historical view has come under intense scrutiny because of growing awareness that in salt-sensitive subjects, acute salt loading does not usually induce greater increases in sodium balance and cardiac output than those induced by salt loading in salt-resistant normotensive controls. Here we highlight pioneering studies from Japan that challenge the historical thinking and provide insights into a contemporary theory of salt sensitivity termed the "vasodysfunction theory." According to this theory, initiation of salt-induced hypertension usually involves abnormal vascular resistance responses to increased salt intake, not greater renal retention of a salt load in salt-sensitive subjects than in normal subjects. By shifting the focus from the historical theory to a contemporary final common pathway for the pathogenesis of salt sensitivity, research from Japan is building the scientific foundation for more effective approaches to the prevention and treatment of salt-induced hypertension. Among the most promising approaches are dietary strategies for reducing the risk for salt-induced hypertension that do not depend on reducing salt consumption in the population.
- MeSH
- arginin analogy a deriváty metabolismus MeSH
- cévní rezistence * MeSH
- dieta MeSH
- hypertenze etiologie prevence a kontrola MeSH
- kuchyňská sůl škodlivé účinky MeSH
- lidé MeSH
- oxid dusnatý metabolismus MeSH
- renální oběh * MeSH
- sodík moč MeSH
- solný stres MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Research Support, N.I.H., Extramural MeSH
The fate of messenger RNA in cytoplasm plays a crucial role in various cellular processes. However, the mechanisms that decide whether mRNA will be translated, degraded or stored remain unclear. Single stranded nucleic acid binding protein (Sbp1), an Arginine-Glycine-Glycine (RGG-motif) protein, is known to promote transition of mRNA into a repressed state by binding eukaryotic translation initiation factor 4G1 (eIF4G1) and to promote mRNA decapping, perhaps by modulation of Dcp1/2 activity. Sbp1 is known to be methylated on arginine residues in RGG-motif; however, the functional relevance of this modification in vivo remains unknown. Here, we report that Sbp1 is arginine-methylated in an hnRNP methyl transferase (Hmt1)-dependent manner and that methylation is enhanced upon glucose deprivation. Characterization of an arginine-methylation-defective (AMD) mutant provided evidence that methylation affects Sbp1 function in vivo. The AMD mutant is compromised in causing growth defect upon overexpression, and the mutant is defective in both localizing to and inducing granule formation. Importantly, the Sbp1-eIF4G1 interaction is compromised both for the AMD mutant and in the absence of Hmt1. Upon overexpression, wild-type Sbp1 increases localization of another RGG motif containing protein, Scd6 (suppressor of clathrin deficiency) to granules; however, this property of Sbp1 is compromised in the AMD mutant and in the absence of Hmt1, indicating that Sbp1 repression activity could involve other RGG-motif translation repressors. Additionally, the AMD mutant fails to increase localization of the decapping activator DEAD box helicase homolog to foci and fails to rescue the decapping defect of a dcp1-2Δski8 strain, highlighting the role of Sbp1 methylation in decapping. Taken together, these results suggest that arginine methylation modulates Sbp1 role in mRNA fate determination.
- MeSH
- aminokyselinové motivy MeSH
- arginin metabolismus MeSH
- cirkulární dichroismus MeSH
- cytoplazmatická granula metabolismus MeSH
- messenger RNA metabolismus MeSH
- metylace MeSH
- posttranslační úpravy proteinů MeSH
- proteiny vázající selen metabolismus MeSH
- Saccharomyces cerevisiae - proteiny metabolismus MeSH
- sekvence aminokyselin MeSH
- vazba proteinů MeSH
- western blotting MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Translation is controlled by numerous accessory proteins and translation factors. In the yeast Saccharomyces cerevisiae, translation elongation requires an essential elongation factor, the ABCF ATPase eEF3. A closely related protein, New1, is encoded by a non-essential gene with cold sensitivity and ribosome assembly defect knock-out phenotypes. Since the exact molecular function of New1 is unknown, it is unclear if the ribosome assembly defect is direct, i.e. New1 is a bona fide assembly factor, or indirect, for instance due to a defect in protein synthesis. To investigate this, we employed yeast genetics, cryo-electron microscopy (cryo-EM) and ribosome profiling (Ribo-Seq) to interrogate the molecular function of New1. Overexpression of New1 rescues the inviability of a yeast strain lacking the otherwise strictly essential translation factor eEF3. The structure of the ATPase-deficient (EQ2) New1 mutant locked on the 80S ribosome reveals that New1 binds analogously to the ribosome as eEF3. Finally, Ribo-Seq analysis revealed that loss of New1 leads to ribosome queuing upstream of 3'-terminal lysine and arginine codons, including those genes encoding proteins of the cytoplasmic translational machinery. Our results suggest that New1 is a translation factor that fine-tunes the efficiency of translation termination or ribosome recycling.
- MeSH
- ABC transportéry chemie genetika metabolismus MeSH
- arginin metabolismus MeSH
- delece genu MeSH
- Escherichia coli genetika metabolismus MeSH
- exprese genu MeSH
- genetické vektory chemie metabolismus MeSH
- interakční proteinové domény a motivy MeSH
- klonování DNA MeSH
- kodon chemie metabolismus MeSH
- konformace proteinů, alfa-helix MeSH
- konformace proteinů, beta-řetězec MeSH
- lysin metabolismus MeSH
- molekulární modely MeSH
- priony chemie genetika metabolismus MeSH
- regulace genové exprese u hub * MeSH
- rekombinantní proteiny chemie genetika metabolismus MeSH
- ribozomy genetika metabolismus MeSH
- Saccharomyces cerevisiae - proteiny chemie genetika metabolismus MeSH
- Saccharomyces cerevisiae genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- sekvenční seřazení MeSH
- terminace translace peptidového řetězce * MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Arteriální hypertenze patří k nejčastějším kardiovaskulárním onemocněním v dospělé populaci. I přes pokroky ve farmakoterapii se nám nedaří dosáhnout cílových hodnot, a tím kontroly krevního tlaku. Léčbu lze zahájit monoterapií nebo kombinací dvou léků v závislosti na klinické situaci. Monoterapie hypertenze bývá úspěšná maximálně u 20–30 % pacientů, u ostatních potřebujeme k dosažení normálních hodnot krevního tlaku kombinaci dvou a více antihypertenziv. Hlavní příčinou neúspěšné normalizace hodnot krevního tlaku je na jedné straně nedostatečné využívání kombinované léčby, především fixních kombinací, a na straně druhé nedostatečná compliance pacientů. Kombinovaná léčba je v současné době považována za nejvýhodnější strategii v léčbě hypertenze.
Arterial hypertension is one of the most common cardiovascular diseases affecting adult population. In spite of advances in pharmacotherapy we continue to fail in attaining target values, and by implication, blood pressure control. Therapy may be initiated with monotherapy or with a combination of two drugs depending on the clinical situation. Monotherapy is usually successful in 20-30% patients with hypertension at the most, while in other patients we need to combine two or more antihypertensive drugs to reach normal blood pressure values. The main cause of our failure to successfully normalize blood pressure values is, on the one hand, inadequate use of combination therapy, especially fixed-dose combinations, and inadequate patient compliance on the other. Combination therapy is currently considered to be the most beneficial strategy in the therapy of hypertension.
- Klíčová slova
- Prestance (perindopril + amlodipin),
- MeSH
- adherence k farmakoterapii MeSH
- antihypertenziva * farmakologie metabolismus terapeutické užití MeSH
- arginin farmakologie metabolismus terapeutické užití MeSH
- blokátory kalciových kanálů terapeutické užití MeSH
- blokátory receptoru 1 pro angiotenzin II terapeutické užití MeSH
- diuretika terapeutické užití MeSH
- fixní kombinace léků MeSH
- hypertenze farmakoterapie MeSH
- inhibitory ACE terapeutické užití MeSH
- kombinace léků amlodipin a olmesartan-medoxomil MeSH
- kombinace léků amlodipin a valsartan MeSH
- lidé MeSH
- perindopril farmakologie metabolismus terapeutické užití MeSH
- Check Tag
- lidé MeSH
Arginine-rich cell-penetrating peptides do not enter cells by directly passing through a lipid membrane; they instead passively enter vesicles and live cells by inducing membrane multilamellarity and fusion. The molecular picture of this penetration mode, which differs qualitatively from the previously proposed direct mechanism, is provided by molecular dynamics simulations. The kinetics of vesicle agglomeration and fusion by an iconic cell-penetrating peptide-nonaarginine-are documented via real-time fluorescence techniques, while the induction of multilamellar phases in vesicles and live cells is demonstrated by a combination of electron and fluorescence microscopies. This concert of experiments and simulations reveals that the identified passive cell penetration mechanism bears analogy to vesicle fusion induced by calcium ions, indicating that the two processes may share a common mechanistic origin.
- MeSH
- arginin metabolismus fyziologie MeSH
- biologický transport MeSH
- buněčná membrána metabolismus MeSH
- fúze membrán účinky léků fyziologie MeSH
- kinetika MeSH
- lipidové dvojvrstvy chemie MeSH
- membrány metabolismus MeSH
- penetrační peptidy chemie metabolismus MeSH
- peptidy chemie fyziologie MeSH
- pseudopodia metabolismus fyziologie MeSH
- simulace molekulární dynamiky MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
