Simultaneous determination of <i>S</i>‐adenosylmethionine and <i>S</i>‐adenosylhomocysteine by capillary zone electrophoresis

K, Pañak; Giorgieri, Sergio A.; Díaz, Luis Eduardo; Ruíz, Oscar Adolfo

doi:10.1002/elps.1150181128

Cited by 14 publications

(15 citation statements)

References 13 publications

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“…Determination of AdoMet and AdoHcy was performed by capillary electrophoresis by using a Waters Capillary Ion Analyzer with an Accusep fused silica gel column (60-cm total length and 75-m i.d.) (15). The retention times of AdoMet and AdoHcy were 8.1 and 12 min, respectively.…”

Section: Construction Of Plasmidsmentioning

confidence: 89%

Involvement of S -adenosylmethionine in G ₁ cell-cycle regulation in Saccharomyces cerevisiae

Mizunuma

Miyamura

Hirata

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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S-adenosyl-L-methionine (AdoMet) is a molecule central to general metabolism, serving as a principal methyl donor for methylation of various cellular constituents. The alteration in the availability of AdoMet has profound effect on cell growth. A mutant allele of Saccharomyces cerevisiae gene SAH1 encoding S-adenosyl-L-homocysteine (AdoHcy) hydrolase, was isolated as a mutation that suppressed the Ca 2؉ -sensitive phenotypes of the zds1⌬ strain, such as the Ca 2؉ -induced, Swe1p-and Cln2p-mediated G 2 cell-cycle arrest, and polarized bud growth. The mutation (sah1-1) led the cells to accumulate AdoMet besides AdoHcy, the substrate of Sah1p. The cells treated with exogenous AdoMet and AdoHcy had markedly decreased levels of SWE1 and CLN2 mRNA, providing the basis for the suppression of the Ca 2؉ sensitivity by the sah1-1 mutation. Exogenous AdoMet transiently led the cells to G1 cellcycle delay whereas AdoHcy caused growth inhibition irrelevant to the cell cycle. The effect of AdoMet in inducing the cell-cycle delay was exerted in a manner independent of Met4p, an overall transcriptional activator for MET genes. Our observation provides an insight into the role played by AdoMet in cell cycle regulation. E ukaryotic cells coordinate cell growth in response to a variety of external signals. In the budding yeast Saccharomyces cerevisiae, commitment to a new round of cell duplication occurs at a control point called Start. Execution of Start demands a sufficient level of G 1 cyclin͞Cdc28 protein kinase activity and is a requirement for DNA synthesis, bud formation, and replication of the spindle pole body (1).Unger and Hertwell (2) found that sulfate starvation of a prototroph, methionine starvation of an auxotroph, or a shift of a conditional methionyl-tRNA synthetase mutant to a restrictive condition led to arrest in G 1 . They proposed that the signal for the nutrients was generated at the level of protein biosynthesis. It was recently shown that disruption of the gene for MET30, an E3 ubiquitin ligase, caused the arrest of G 1 (3). MET30 is an essential gene that encodes a negative regulator of the sulfur amino acid biosynthesis pathway (4, 5). The uncontrolled accumulation of Met4p, a transcriptional activator for MET genes (6), in a met30⌬ strain has been suggested to inhibit passage of Start by preventing the accumulation of G 1 transcripts, such as CLN1 and CLN2 (3). It is therefore reasonable to expect some sort of connection between the cyclin-Cdc28p kinase complex and several external signals, including nutrient signals, although the underlying mechanisms are not completely known (7).L-methionine is an essential amino acid required for protein synthesis and participates, together with ATP, in the formation of S-adenosylmethionine (AdoMet). AdoMet is a high-energy sulfonium compound that is used in the majority of biological methylation reactions (7, 8). On demethylation, AdoMet is converted into S-adenosylhomocysteine (AdoHcy), a molecule that can be further hydrolyzed to adenosine and homocysteine by AdoH...

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Section: Construction Of Plasmidsmentioning

confidence: 89%

Involvement of S -adenosylmethionine in G ₁ cell-cycle regulation in Saccharomyces cerevisiae

Mizunuma

Miyamura

Hirata

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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“…The hepatic SAM level was quantified by the capillary electrophoresis method of Panak. 21) In brief, frozen liver was homogenized in 5 volumes (v/wt) of ice-cold 4% perchloric acid solution. The homogenate was centrifuged at 13;000 Â g for 10 min at 4 C, and the supernatant was used for measurement after the appropriate dilution.…”

Section: Methodsmentioning

confidence: 99%

Sake Yeast Suppresses Acute Alcohol-Induced Liver Injury in Mice

Izu¹,

Shobayashi²,

Manabe³

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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Brewer's and baker's yeasts appear to have components that protect from liver injury. Whether sake yeast, Saccharomyces cerevisiae Kyokai no. 9, also has a hepatoprotective effect has not been examined. Here we show that sake yeast suppresses acute alcoholic liver injury in mice. Male C57BL/6 mice that had been fed a diet containing 1% sake yeast for two weeks received three doses of ethanol (5 g/kg BW). In the mice fed sake yeast, ethanol-induced increases in triglyceride (TG) and glutamate pyruvate transaminase (GPT) were significantly attenuated and hepatic steatosis was improved. In addition, sake yeast-fed mice showed a smaller decrease in hepatic S-adenosylmethionine (SAM) level and a smaller increase in plasma homocysteine (Hcy) level after ethanol treatment than the control mice, suggesting that a disorder of methonine metabolism in the liver caused by ethanol was relieved by sake yeast. These results indicate that sake yeast protects against alcoholic liver injury through maintenance of methionine metabolism in the liver.Key words: Saccharomyces cerevisiae; sake yeast; liver injury; alcohol; methionine metabolismThe physiological effects of brewer's yeast and its components in mammals have been well studied. For example, brewer's yeast cell wall reduces serum lipid levels in rats, 1) and improves defecation by contributing to the fermentation ability, water holding capacity, and swelling force of the large intestine.2) To understand the gene expression of Saccharomyces bayanuis as a lager brewer's yeast during beer fermentation, sequencing of the whole genome was performed.3) This provided further information about the physiological effects of brewer's yeast as well as the brewing of beer.On the other hand, there have been few studies of the physiological effects of sake yeast, Saccharomyces cerevisiae Kyokai yeast, which is used in sake (Japanese rice wine) brewing. Manabe et al. reported that both sake yeast and sake lees, the leftovers of sake brewing, increase spontaneous locomotive activity in rats. 4) They proposed that the effect of sake lees partly depends on sake yeast. The sake lees contain rice components that are not assimilated by sake yeast and sake koji, Aspergillus oryzae, components of sake yeast and sake koji, and their metabolites. Therefore, to understand the physiological effects of sake yeast may help to understand the physiological effcts of sake lees and to find new uses for them.A distinctive characteristic of sake yeast is that it is able to accumulate higher levels of S-adenosylmethionine (SAM) intracellularly than other yeasts, bacteria, molds, and some other microorganisms.5,6) SAM functions as a major methyl group donor in transmethylation of proteins, nucleic acids, polysaccharides, phospholipids, and fatty acids and as a precursor of glutathione. It has been proposed as a chemotherapeutic agent for alcoholic liver disease, 7) depression, 8) osteoarthritis , 9) and Alzheimer's disease. 10) Another distinctive characteristic of sake yeast is that it has a higher ethanol t...

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“…In the oldest one [9], SAM and SAH were separated in 40 mM phosphate buffer, pH 2.50, as BGE, in uncoated capillaries, in a run time shorter than 8 min, with a mass detection limit in the range of 10 fmol. In the second report [10], SAM and SAH were also separated by CZE in uncoated capillaries in a BGE of 200 mM Gly-HCl, pH 1.8, but the analysis was not performed on purified, commercial products, instead in mouse and rat tissues.…”

Section: Introductionmentioning

confidence: 99%

Analysis of trace degradation products (decarboxylated diastereoisomers) of S‐adenosylmethionine by electrophoresis in capillaries with cationic coatings (N‐methylpolyvinylpyridinium or divalent barium)

et al. 2010

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Commercial preparations of S-adenosylmethionine (SAM) when analyzed in uncoated capillaries show a minute impurity believed to be decarboxylated (dc) SAM. By using two types of cationic coatings, thus reducing the electro-endo-osmotic flow (EOF), it was possible to separate this impurity into two diastereoisomers of dcSAM. The coatings evaluated for this purpose were: (i) N-methylpolyvinylpyridinium, used under reversed EOF at acidic conditions (pH 4.0) and (ii) deposition of divalent barium at alkaline pH values (pH 9.4), providing reduced EOF. Under these conditions, it was possible to separate this impurity into two diastereoisomers, which by chemical synthesis were indeed proven to be dcSAM. It was further demonstrated that, in the alkylation of 5'-methylthioadenosine by 3-bromopropylamine in bromidric acid to dcSAM, another minute impurity was present, proven, via mass spectrometry, to consist of S-(5'-adenosyl)-3-thiopropylamine (decarboxylated and demethylated (dc-SAH)). The LOD for the two dcSAM diastereoisomers was assessed as 17.5 μg/mL and their LOQ as 25.5 μg/mL. By the barium-based protocol it was possible to quantify the dcSAM, present in a commercial sample of SAM, as a 0.1% impurity.

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Simultaneous determination of S‐adenosylmethionine and S‐adenosylhomocysteine by capillary zone electrophoresis

Cited by 14 publications

References 13 publications

Involvement of S -adenosylmethionine in G ₁ cell-cycle regulation in Saccharomyces cerevisiae

Involvement of S -adenosylmethionine in G ₁ cell-cycle regulation in Saccharomyces cerevisiae

Sake Yeast Suppresses Acute Alcohol-Induced Liver Injury in Mice

Analysis of trace degradation products (decarboxylated diastereoisomers) of S‐adenosylmethionine by electrophoresis in capillaries with cationic coatings (N‐methylpolyvinylpyridinium or divalent barium)

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Simultaneous determination of S‐adenosylmethionine and S‐adenosylhomocysteine by capillary zone electrophoresis

Cited by 14 publications

References 13 publications

Involvement of S -adenosylmethionine in G 1 cell-cycle regulation in Saccharomyces cerevisiae

Involvement of S -adenosylmethionine in G 1 cell-cycle regulation in Saccharomyces cerevisiae

Sake Yeast Suppresses Acute Alcohol-Induced Liver Injury in Mice

Analysis of trace degradation products (decarboxylated diastereoisomers) of S‐adenosylmethionine by electrophoresis in capillaries with cationic coatings (N‐methylpolyvinylpyridinium or divalent barium)

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Involvement of S -adenosylmethionine in G ₁ cell-cycle regulation in Saccharomyces cerevisiae

Involvement of S -adenosylmethionine in G ₁ cell-cycle regulation in Saccharomyces cerevisiae