287 research outputs found
A resonance Raman study of the binding of ethanol and methanol to ferrihemoglobin
AbstractThe interactions of ethanol and methanol with ferrihemoglobin were examined using resonance Raman spectroscopy. After binding either alcohol, the low-frequency resonance Raman spectra of human ferrihemoglobin are almost identical to the unperturbed spectrum except for shifts in the 309 cmâ1 band to higher frequency by as much as 8 cmâ1. The ethanol-induced shift is greater than that with methanol even though complex formation was less for ethanol than methanol. The spectral changes imply a site-specific, similar binding of these alcohols to ferrihemoglobin which may involve steric interactions. Possible assignments of the 309 cmâ1 band to structural features as well as potential mechanisms of the alcohol-induced spectral changes are discussed
Bongkrekic acid and atractyloside inhibits chloride channels from mitochondrial membranes of rat heart
AbstractThe aim of this work was to characterize the effect of bongkrekic acid (BKA), atractyloside (ATR) and carboxyatractyloside (CAT) on single channel properties of chloride channels from mitochondria. Mitochondrial membranes isolated from a rat heart muscle were incorporated into a bilayer lipid membrane (BLM) and single chloride channel currents were measured in 250/50Â mM KCl cis/trans solutions. BKA (1â100Â ÎźM), ATR and CAT (5â100Â ÎźM) inhibited the chloride channels in dose-dependent manner. The inhibitory effect of the BKA, ATR and CAT was pronounced from the trans side of a BLM and it increased with time and at negative voltages (transâcis). These compounds did not influence the single channel amplitude, but decreased open dwell time of channels. The inhibitory effect of BKA, ATR and CAT on the mitochondrial chloride channel may help to explain some of their cellular and/or subcellular effects
Inorganic Reactive Sulfur-Nitrogen Species: Intricate Release Mechanisms or Cacophony in Yellow, Blue and Red?
Since the heydays of Reactive Sulfur Species (RSS) research during the first decade of the Millennium, numerous sulfur species involved in cellular regulation and signalling have been discovered. Yet despite the general predominance of organic species in organisms, recent years have also seen the emergence of inorganic reactive sulfur species, ranging from inorganic polysulfides (HSxâ/Sx2â) to thionitrous acid (HSNO) and nitrosopersulfide (SSNOâ). These inorganic species engage in a complex interplay of reactions in vitro and possibly also in vivo. Employing a combination of spectrophotometry and sulfide assays, we have investigated the role of polysulfanes from garlic during the release of nitric oxide (â˘NO) from S-nitrosoglutathione (GSNO) in the absence and presence of thiol reducing agents. Our studies reveal a distinct enhancement of GSNO decomposition by compounds such as diallyltrisulfane, which is most pronounced in the presence of cysteine and glutathione and presumably proceeds via the initial release of an inorganic mono- or polysulfides, i.e., hydrogen sulfide (H2S) or HSxâ, from the organic polysulfane. Albeit being of a preliminary nature, our spectrophotometric data also reveals a complicated underlying mechanism which appears to involve transient species such as SSNOâ. Eventually, more in depth studies are required to further explore the underlying chemistry and wider biological and nutritional implications of this interplay between edible garlic compounds, reductive activation, inorganic polysulfides and their interplay with â˘NO storage and release
Inorganic Polysulfides and Related Reactive SulfurâSelenium Species from the Perspective of Chemistry
Polysulfides (H2Sx) represent a class of reactive sulfur species (RSS) which includes
molecules such as H2S2, H2S3, H2S4, and H2S5, and whose presence and impact in biological systems,
when compared to other sulfur compounds, has only recently attracted the wider attention of
researchers. Studies in this field have revealed a facet-rich chemistry and biological activity associated
with such chemically simple, still unusual inorganic molecules. Despite their chemical simplicity,
these inorganic species, as reductants and oxidants, metal binders, surfactant-like âcork screwsâ for
membranes, components of perthiol signalling and reservoirs for inorganic hydrogen sulfide (H2S),
are at the centre of complicated formation and transformation pathways which affect numerous
cellular processes. Starting from their chemistry, the hidden presence and various roles of polysulfides
in biology may become more apparent, despite their lack of clear analytical fingerprints and often
murky biochemical footprints. Indeed, the biological chemistry of H2Sx follows many unexplored
paths and today, the relationship between H2S and its oxidized H2Sx species needs to be clarified as
a matter of âunmistaken identityâ. Simultaneously, emerging species, such as HSSeSH and SenS8ân,
also need to be considered in earnest
EPR Study of KO2 as a Source of Superoxide and â˘BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H2S and Se-Derivatives
: Superoxide radical anion (O2
â˘â) and its derivatives regulate numerous physiological and
pathological processes, which are extensively studied. The aim of our work was to utilize KO2 as a
source of O2
â˘â and the electron paramagnetic resonance (EPR) spin trapping 5-tert-butoxycarbonyl-5-
methyl-1-pyrroline N-oxide (BMPO) technique for the preparation of â˘BMPO-OOH and/or â˘BMPOOH radicals in water solution without DMSO. The method distinguishes the interactions of various
compounds with â˘BMPO-OOH and/or â˘BMPO-OH radicals over time. Here, we show that the
addition of a buffered BMPO-HCl mixture to powdered KO2
formed relatively stable â˘BMPO-OOH
and â˘BMPO-OH radicals and H2O2
, where the â˘BMPO-OOH/OH ratio depended on the pH. At a
final pH of ~6.5â8.0, the concentration of â˘BMPO-OOH radicals was âĽ20 times higher than that of
â˘BMPO-OH, whereas at pH 9.0â10.0, the â˘BMPO-OH radicals prevailed. The â˘BMPO-OOH/OH
radicals effectively cleaved the plasmid DNA. H2S decreased the concentration of â˘BMPO-OOH/OH
radicals, whereas the selenium derivatives 1-methyl-4-(3-(phenylselanyl) propyl) piperazine and
1-methyl-4-(4-(phenylselanyl) butyl) piperazine increased the proportion of â˘BMPO-OH over the
â˘BMPO-OOH radicals. In conclusion, the presented approach of using KO2 as a source of O2
â˘â/H2O2
and EPR spin trap BMPO for the preparation of â˘BMPO-OOH/OH radicals in a physiological solution
could be useful to study the biological effects of radicals and their interactions with compounds
The reaction products of sulfide and S-nitrosoglutathione are potent vasorelaxants
The chemical interaction of sodium sulfide (Na2S) with the NO-donor S-nitrosoglutathione (GSNO) has been described to generate new reaction products, including polysulfides and nitrosopersulfide (SSNO-) via intermediacy of thionitrous acid (HSNO). The aim of the present work was to investigate the vascular effects of the longer-lived products of the Sulfide/GSNO interaction. Here we show that the products of this reaction relax precontracted isolated rings of rat thoracic aorta and mesenteric artery (but to a lesser degree rat uterus) with a >2-fold potency compared with the starting material, GSNO (50?nM), whereas Na2S and polysulfides have little effect at 1-5?ÂľM. The onset of vasorelaxation of the reaction products was 7-10 times faster in aorta and mesenteric arteries compared with GSNO. Relaxation to GSNO (100-500?nM) was blocked by an inhibitor of soluble guanylyl cyclase, ODQ (0.1 and 10?ÂľM), and by the NO scavenger cPTIO (100?ÂľM), but less affected by prior acidification (pH 2-4), and unaffected by N-acetylcysteine (1?mM) or methemoglobin (20?ÂľM heme). By contrast, relaxation to the Sulfide/GSNO reaction products (100-500?nM based on the starting material) was inhibited to a lesser extent by ODQ, only slightly decreased by cPTIO, more markedly inhibited by methemoglobin and N-acetylcysteine, and abolished by acidification before addition to the organ bath. The reaction mixture was found to generate NO as detected by EPR spectroscopy using N-(dithiocarboxy)-N-methyl-D-glucamine (MGD2)-Fe2+ as spin trap. In conclusion, the Sufide/GSNO reaction products are faster and more pronounced vasorelaxants than GSNO itself. We conclude that in addition to NO formation from SSNO-, reaction products other than polysulfides may give rise to nitroxyl (HNO) and be involved in the pronounced relaxation induced by the Sulfide/GSNO cross-talk
The β Subunit Increases the Ca2+ Sensitivity of Large Conductance Ca2+-activated Potassium Channels by Retaining the Gating in the Bursting States
Coexpression of the β subunit (KV,Caβ) with the Îą subunit of mammalian large conductance Ca2+- activated K+ (BK) channels greatly increases the apparent Ca2+ sensitivity of the channel. Using single-channel analysis to investigate the mechanism for this increase, we found that the β subunit increased open probability (Po) by increasing burst duration 20â100-fold, while having little effect on the durations of the gaps (closed intervals) between bursts or on the numbers of detected open and closed states entered during gating. The effect of the β subunit was not equivalent to raising intracellular Ca2+ in the absence of the beta subunit, suggesting that the β subunit does not act by increasing all the Ca2+ binding rates proportionally. The β subunit also inhibited transitions to subconductance levels. It is the retention of the BK channel in the bursting states by the β subunit that increases the apparent Ca2+ sensitivity of the channel. In the presence of the β subunit, each burst of openings is greatly amplified in duration through increases in both the numbers of openings per burst and in the mean open times. Native BK channels from cultured rat skeletal muscle were found to have bursting kinetics similar to channels expressed from alpha subunits alone
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Nitrosopersulfide (SSNO-) targets soluble guanylyl cyclase and induces vasodilation in vivo
Background
Recent experimental evidence suggests that nitric oxide (NO) and hydrogen sulfide signaling pathways are intimately intertwined particularly in the vasculature, with mutual attenuation or potentiation of biological responses under control of the soluble guanylyl cyclase (sGC) / phopshodiesterase (PDE) pathway. There is now compelling evidence that part of the NO/sulfide cross talk has a chemical foundation via the formation of S/N-hybrid molecules including thionitrous acid (HSNO) and nitrosopersulfde (SSNO-). The aim of this study was to characterize the bioactive products of the interaction between sulfide and NO metabolites targeting sGC that may potentially regulate vasodilation.
Results
We found that the chemical interaction of sulfide with NO or nitrosothiols leads to formation of S/N-hybrid metabolites including SSNO- via intermediate formation of HSNO. Contrary to a recent report in the literature but consistent with the transient nature of HSNO, its formation was not detectable by high-resolution mass spectrometry under physiologically relevant conditions. SSNO- is also formed in non-aqueous media by the reaction of nitrite with oxidized sulfur species including colloidal sulfur and polysulfides. SSNO- is stable in the presence of high concentrations of thiols, release NO, and activates sGC in RFL-6 cells in an NO-dependent fashion. Moreover, SSNO- is a potent vasodilator in aortic rings in vitro and lowers blood pressure in rats in vivo. The presence of high concentrations of SOD or thiols does not affect SSNO- mediated sGC activation, while it potentiates and inhibits the effects of the nitroxyl (HNO) donor Angeli's salt, suggesting that HNO release from SSNO- is not involved in sGC activation.
Conclusion
The reaction between NO and sulfide leads to fomation of S/N-hybrid molecules including SSNO-, releasing NO, activating sGC and inducing vasodilation. SSNO- is considerably more stable than HSNO at pH 7.4 and thus a more likely biological mediator that can account for the chemical cross-talk between NO and sulfide
Doxorubicin Causes Lesions in the Electron Transport System of Skeletal Muscle Mitochondria that are Associated with a Loss of Contractile Function
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