Cysteine Sulfinate Research Articles

Expression and clinical significance of CSAD in type I endometrial carcinoma

The current study aimed to identify the expression and clinical significance of cysteine sulfinate decarboxylase (CSAD) in type I endometrial carcinoma (EC). A total of 350 patients with endometrial lesions were involved in the study at Zhejiang University’s Women’s Hospital. Immunochemical staining was performed on samples of normal endometrium (NE), atypical endometrial hyperplasia (AEH), and type I EC. The study evaluated the association of CSAD staining with clinical characteristics and patient survival. CSAD expression in type I EC was elevated in comparison with NE, and increased expression correlates with International Federation of Gynecology and Obstetrics (FIGO) stages, Lymph vascular space invasion (LVSI), and Lymph node metastasis (LNM). High CSAD levels are associated with poor disease-free survival and overall survival. small interfering RNAs (siRNAs) can decrease CSAD expression, inhibit cancer cell proliferation, and affect B-cell lymphoma-2 (BCL-2) and BCL2-Associated X (BAX) protein expression. Our research indicates that higher levels of CSAD expression in type I EC was significantly associated with some clinical variables indicating poor prognosis and survival rate in patients with type I EC. patients and could promote cancer cell growth by affecting cell apoptosis rate.

Identification and expression of cysteine sulfite decarboxylase genes and their response to hyposalinity stress in Ruditapes philippinarum

Cysteine sulfinate decarboxylase (CSAD) is a rate-limiting enzyme in taurine biosynthesis. In this study, the structure and expression characteristics of the RpCSAD gene in Ruditapes philippinarum were analyzed. The pyridoxal phosphate (PLP)-dependent amino acid carboxylase structural domains and catalytic active sites of six copies of RpCSAD were highly conserved. Tissue expression analysis demonstrated that RpCSAD1 and RpCSAD3-6 was primarily expressed in the mantle tissue, and RpCSAD2 in the water tube and gonad tissue. After exposure to hyposalinity stress, the expression levels of RpCSADs were found to be upregulated in all three shell-colored groups of clams. Remarkably, there was a pronounced increase in the expression of RpCSAD1-6 observed specifically in the zebra shell-color groups. These findings strongly suggest the involvement of taurine metabolism in the response of zebra shell-color clams to low salinity stress, thereby emphasizing its critical role in the adaptation mechanisms. These findings could contribute to a better understanding the mechanisms of osmotic stress tolerance in shellfish.

МЕТАБОЛИЧЕСКИЙ ПРОФИЛЬ СВОБОДНЫХ АМИНОКИСЛОТ ПРИ БЕРЕМЕННОСТИ, ОСЛОЖНЕННОЙ COVID-19

Background. The unexpected emergence and rapid spread around the world of a new coronavirus infection COVID-19, with a large number of deaths, has shown the importance of developing the methods for predicting a severe course of viral infections, creating diagnostic tests, preventive vaccines and drugs that act on key factors in the course and progression of the disease. Clinical studies examining metabolomic changes in pregnant women with COVID-19 infection are virtually non-existent. Purpose of the study. To study the features of the metabolic profile of free amino acids during pregnancy complicated by COVID-19. Material and methods. The concentrations of free amino acids and their derivatives in the blood plasma of 86 women were studied. The main group included 51 pregnant women with a confirmed diagnosis of COVID-19; the control group was composed of 35 somatically healthy women with uncomplicated pregnancies. The analysis of amino acids and their derivatives was carried out using high performance liquid chromatography. Results. During the development of COVID-19 infection, protein catabolism predominates, as evidenced by an increase in the concentration of 28 amino acids and their derivatives in venous blood plasma. We revealed a statistically significant increase in the concentration of cysteine, homocysteine, γ-glutamylcysteine, phosphoserine, cysteine sulfinate, aspartate, homocysteate, glutamate, asparagine, serine, threonine, 1-methylhistidine, glycine, citrulline, arginine, alanine, symmetric dimethylarginine, asymmetric dimethylarginine, β-aminobutyric acid, γ-aminobutyric acid, tyrosine, valine, methionine, phenylalanine, leucine, isoleucine, lysine, ornithine in pregnant women with a new coronavirus infection as compared to the patients in the control group. Conclusion. The study of the features of the metabolic profile of free amino acids in pregnant women with a new coronavirus infection will enable us to identify additional biomarkers of the severity of the disease and predict the course of the disease in patients during the gestation period.

The Effect of Dietary Cysteine Level on Cysteine Metabolism in Rats

Male, Sprague-Dawley rats were fed l-amino acid diets that contained 0.4% l methionine and either 0, 0.2% (control), or 2.6% l-cysteine (free base) for 5 or 20 days. Hepatic cysteine dioxygenase activity in rats fed 2.6% cysteine was 5 and 3-times as great as in pair-fed control rats at 5 and 20 days, respectively. Cysteine sulfinate decarboxylase activity in liver of rats fed 2.6% cysteine for 20 days was about 40% of the pair-fed control level. The activity of cysteine sulfinate aminotransferase and the rate of cysteine desulfhydration were not influenced by dietary cysteine content. Rats that had been fed these diets for 5 days were intubated with 5 g of diet that contained either 0.2% or 2.6% l-[35S]cysteine. The 24-hour urinary excretion of total 35S, 35SO4, and [35S]taurine by rats that had been fed 2.6% cysteine for 5 days and intubated with 0.2% l-[35S]cysteine was 1.4-, 1.8-, and 2.4-fold, respectively, that of pair-fed control rats. About 2.1, 1.8, and 9.1 times as much 35S, 35SO/4, and [35S]taurine, respectively, were excreted by rats fed 2.6% cysteine for 5 days and intubated with 2.6% l-[35S]cysteine as was excreted by pair-fed control rats. Urinary excretion of these metabolites was not different between rats fed 0 and 0.2% cysteine. These results indicate that the cysteine sulfinate pathway plays a major regulatory role in the metabolism of excess cysteine.cysteine cysteine desulfhydrase cysteine dioxygenase cysteine sulfinate decarboxylase taurine sulfate

Heterologous expression and functional characterization of cysteamine dioxygenase from the deep-sea mussel Bathymodiolus septemdierum

In invertebrates inhabiting hydrothermal vent areas, hypotaurine, a precursor of taurine, is thought to mitigate the toxicity of hydrogen sulfide in vent fluids. Information about hypotaurine synthesis pathways in invertebrates is limited, although two pathways, the cysteamine [2-aminoethanethiol (AET)] pathway and the cysteine sulfinate (CSA) pathway are known in mammals. In this study, we cloned a cDNA encoding AET dioxygenase (ADO), the central enzyme of the AET pathway, from the vent mussel Bathymodiolus septemdierum. In the encoded protein (BsADO), functionally important residues, including metal-binding histidines, are conserved. In maximum likelihood phylogenetic analysis, BsADO clustered with ADOs of other invertebrates. By reverse transcription PCR, BsADO mRNA was detected in all tissues examined at similar levels, suggesting that its function is distinct from that of the CSA pathway, predominantly expressed in the gill. BsADO with a His tag, expressed in Escherichia coli in the presence of Fe2+, converted AET to hypotaurine, but BsADO expressed in the absence of iron exhibited lower activity. BsADO was active from pH 8 to 11, and from 0 °C to 37 °C, with a peak at 20 °C. This is the first functional characterization of ADO in marine invertebrates.

Testosterone enhances taurine synthesis by upregulating androgen receptor and cysteine sulfinic acid decarboxylase expressions in male mouse liver.

Taurine is an end-product of cysteine metabolism, whereas cysteine dioxygenase (CDO) and cysteine sulfinate decarboxylase (CSAD) are key enzymes regulating taurine synthesis. Sex steroids, including estrogens and androgens, are associated with liver physiopathological processes; however, we still do not know whether taurine and sex steroids interact in regulating liver physiology and hepatic diseases, and whether there are sex differences, although our recent study shows that the estrogen is involved in regulating taurine synthesis in mouse liver. The present study was thus proposed to identify whether 17-β-estradiol and testosterone (T) play their roles by regulating CDO and CSAD expression and taurine synthesis in male mouse liver. Our results demonstrated that testosterone did not have a significant influence on CDO expression but significantly enhanced CSAD, androgen receptor (AR) expressions, and taurine levels in mouse liver, cultured hepatocytes, and HepG2 cells, whereas these effects were abrogated by AR antagonist flutamide. Furthermore, our results showed that testosterone increased CSAD-promoter-luciferase activity through the direct interaction of the AR DNA binding domain with the CSAD promoter. These findings first demonstrate that testosterone acts as an important factor to regulate sulfur amino acid metabolism and taurine synthesis through AR/CSAD signaling pathway. In addition, the in vivo and in vitro experiments showed that 17-β-estradiol has no significant effects on liver CSAD expression and taurine synthesis in male mice and suggest that the effects of sex steroids on the taurine synthesis in mouse liver have sex differences. These results are crucial for understanding the physiological functions of taurine/androgen and their interacting mechanisms in the liver.NEW & NOTEWORTHY This study demonstrates that testosterone functions to enhance taurine synthesis by interacting with androgen receptor and binding to cysteine sulfinate decarboxylase (CSAD) promoter zone. Whereas estrogen has no significant effects either on liver CSAD expression or taurine synthesis in male mice and suggests that the effects of sex steroids on taurine synthesis in the liver have gender differences. These new findings are the potential for establishing effective protective and therapeutic strategies for liver diseases.

식이제한이 마우스 신장의 글루타치온 대사과정에 미치는 영향

Various benefits for the maintenance of metabolic homeostasis can be obtained through dietary restriction. Short-term fasting of mice may induce systemic metabolic remodeling, which may have a significant effect on the level of endogenous metabolites. This study was conducted to examine metabolic changes of sulfur-containing substances in the kidneys of fasted mice over 48 h. No change in the concentration of renal methionine was observed over 48 h; however, significant decreases in the concentrations of S-adenosylmethionine and S-adenosylhomocysteine were observed at both 24 and 48 h after fasting. A significant decrease in the concentration of renal cysteine was observed from 24 h, and this was maintained at 48 h. A significant decrease in the concentration of glutathione (GSH) was observed in the kidney at 48 h, whereas increased concentrations of taurine were observed 48 h after fasting. Although significantly increased expression levels of BHMT were observed, markedly decreased expression levels of CβS were observed at 48 h after fasting, indicating activated generation of methionine, but downregulation of cysteine synthesis. The expression level of γ-GCL was not altered by fasting for 48 h; however, a significant increase of cysteine sulfinate decarboxylase mediating the taurine synthesis was observed. Significantly decreased protein expression levels of solute carrier family 7 member 11, which imports extracellular cystine, were observed at 48 h. These results suggest that the renal methionine concentration was maintained during short-term fasting through an increase in the re-methylation of homocysteine. Moreover, a reduction in the concentration of cysteine was the result of an increase in the synthesis of taurine and a decrease in the uptake of extracellular cystine, which may result in a decreased concentration of renal GSH.

In vitro and in vivo study on prevention of myocardial ischemic injury by taurine.

BackgroundMyocardial ischemia (MI) often causes angina, arrhythmia, and cardiac insufficiency, sometimes resulting in death. Ischemia-induced myocardial tissue damage is attributed to the hypoxic damage of myocardial cells producing apoptosis and decreased proliferation. Taurine has been shown to improve MI, but its mechanism is largely unknown.MethodsIn this study, the relationship between taurine and severity of MI in vivo was evaluated by quantifying myocardial infarct areas and metabolic indicators of myocardial damage and measuring taurine levels in cardiac muscle and plasma by high performance liquid chromatography (HPLC). To elucidate how taurine might suppress ischemic injury, we established an in vitro ischemia model with isolated primary rat cardiomyocytes cultured without serum or glucose and under hypoxia. We evaluated the indicators of MI and damage, including lactic dehydrogenase (LDH), creatine kinase (CK), and cardiac troponin I (cTnI). We also examined the levels of taurine transporter (TauT), cysteine dioxygenase (CDO), and cysteine sulfinate decarboxylase (CSD) proteins involved in transport and synthesis of taurine in the myocardium and those of 2 apoptosis-associated proteins, namely, Bcl-2 associated X protein (BAX) and B-cell lymphoma-2 (Bcl-2).ResultsExposure of myocardial cells to ischemia led to the decrease of taurine content, the suppression of cell proliferation, and led to calcium ion overload and apoptosis. Pretreatment with taurine alleviated the ischemic damage, with concomitant elevation of intracellular taurine concentrations. Molecular mechanism analysis showed that pretreatment with taurine upregulated the TauT, CDO, and CSD, 2 rate-limiting enzymes involved in taurine synthesis. These effects facilitated both taurine transport into cells and taurine synthesis, leading to taurine accumulation. In addition, apoptosis inhibition by taurine appeared to be mediated by upregulated Bcl-2 and downregulated BAX, as well as inhibition of calcium overload by suppression of calcium binding protein.ConclusionsWe demonstrated that TauT is critical for the attenuation of myocardial ischemic damage by taurine, facilitating taurine absorption and synthesis. These findings provided new insights and a theoretical foundation for future studies examining taurine as a potential treatment for MI.

Changes in the concentration of sulfur-containing amino acids in the brain after methionine load in the experimentChanges in the concentration of sulfur-containing amino acids in the brain after methionine load in the experiment

The effect of methionine overload on the state of the pool of sulfur-containing amino acids and their metabolites was studied in the various brain structures determined by reverse phase high performance liquid chromatography (HPLC). In all regions of the brain studied, methionine led to a unidirectional imbalance of sulfur-containing compounds: there was an increase in the concentrations of methionine, cystathionine and hypotaurine. The most pronounced increase in methionine and hypotaurine levels was observed in the striatum, cystathionine in the hemispheres. A significant increase in taurine concentration was observed only in the hypothalamus and striatum. In other parts of the brain a tendency to increase its level was shown. In all brain regions studied except the striatum, serine levels were decreased. In the cerebellum, in comparison with other regions, an increase in the level of cysteic acid and a decrease in the level of cysteinesulfinic acid were observed, which indicates that taurine synthesis is occurred mainly through the cysteine sulfinate oxidation.

Effects of DL-methionine and a methionine hydroxy analogue (MHA-Ca) on growth, amino acid profiles and the expression of genes related to taurine and protein synthesis in common carp (Cyprinus carpio)

In this trial, nine diets were formulated to investigate the effects of DL-methionine (DL-Met) and a methionine hydroxy analogue (MHA-Ca) on growth, amino acid profiles and the expression of genes related to taurine and protein synthesis in common carp. The basal diet was prepared without methionine supplementation and the other diets were formulated with 0.1%, 0.2%, 0.3% and 0.4% of DL-Met or MHA-Ca, respectively. After the eight-week feeding trial, fish fed with DL-Met or MHA-Ca supplemented diets had higher growth performance and feed utilization when compared with basal diet and two-way ANOVA showed the growth performance and feed utilization were significantly increased in fish fed DL-Met supplemented diets (P < 0.05). The concentrations of free amino acids in serum (Met, Leu, His, Lys, Val, Arg, Ile, Phe and Tau) and muscle (Tau) significantly increased as dietary DL-Met and MHA-Ca increased. Moreover, two-way ANOVA showed that taurine was higher in common carp fed with DL-Met diets compared with MHA-Ca diets. To find out how DL-Met and MHA-Ca affect taurine synthesis, genes involved in methionine degradation and taurine synthesis were further investigated. DL-Met or MHA-Ca supplementation decreased the expression of methionine adenosyltransferase II, alpha (MAT2A) and methionine synthase (MS) and increased the expression of the cysteine dioxygenase (CDO). Moreover, DL-Met supplementation significantly increased the expression of CDO and cysteine sulfinate acid decarboxylase (CSD) when compared with MHA-Ca supplementation. As for protein synthesis, the mRNA expression of mechanistic target of rapamycin (mTOR) in liver was not significantly different among all diet groups. But the expression of eukaryotic translation initiation factor 4E-Binding protein 1 (4EBP1), general control nonderepressible2 (GCN2) and asparagine synthetase (ASNS) in liver were significantly down-regulated as dietary DL-Met or MHA-Ca increased. Compared with MHA-Ca diets, the expression of S6 (ribosomal protein s6) and eIF4E (eukaryotic translation initiation factor 4E binding protein 2) were markedly elevated in DL-Met groups by the analysis of two-way ANOVA. This study indicated that dietary DL-Met or MHA-Ca supplement could increase free essential amino acids in serum, as well as improve taurine synthesis and protein synthesis in common carp. Multi-exponential regression analysis showed that MHA-Ca was less utilized by common carp than DL-Met with bioavailability values of 41% to 50% on weight-for-weight basis.

Dietary taurine supplementation affects lipid metabolism and improves the oxidative status of European seabass (Dicentrarchus labrax) juveniles

The aim of this study was to assess the effect of dietary taurine (Tau) in intermediary metabolism, oxidative status, and Tau biosynthesis capacity in European seabass juveniles. For that purpose, four diets (45% protein and 18% lipid) were formulated with increasing levels of Tau (0.2, 0.5, 0.7, and 1.2% of DM) and fed to triplicate groups of fish (IBW 55g) until apparent visual satiety for 10 weeks.Total bile acid content in the liver increased as dietary Tau supplementation increase, while the opposite occurred in the plasma. Plasma glucose, total cholesterol, HDL, and LDL cholesterol decreased, while total and indirect bilirubin increased with the increase of dietary Tau level.The activity of the key intermediary enzymes measured decreased with the increase of dietary Tau up to 0.7%. Regarding the antioxidant-related enzymes, dietary Tau supplementation decreased hepatic catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase activities with the maximum effects being observed as dietary Tau increased from 0.2 to 0.5%. Overall, there was a decreasing trend of hepatic lipid peroxidation with the increase of dietary Tau.This study is the first to demonstrate that in European seabass juveniles the capacity of expressing cysteine dioxygenase, an enzyme involved in the cysteine sulfinate pathway for Tau synthesis, although its expression was not affected by Tau levels in the diet.This study indicates that dietary Tau levels affect intermediary metabolism, namely carbohydrates and lipidic metabolism, and improve the antioxidant status of European seabass juveniles. Furthermore, although CDO expression was detected, European seabass seems unable to regulate its activity, these results supporting the essentiality of Tau for this species.

Structural preferences of cysteine sulfinic acid: The sulfinate engages in multiple local interactions with the peptide backbone

Cysteine sulfinic acid (Cys-SO2–) is a non-enzymatic oxidative post-translational modification (PTM) that has been identified in hundreds of proteins. However, the effects of cysteine sulfination are in most cases poorly understood. Cys-SO2– is structurally distinctive, with long sulfur-carbon and sulfur-oxygen bonds, and with tetrahedral geometry around sulfur due to its lone pair. Cys-SO2– thus has a unique range of potential interactions with the protein backbone which could facilitate protein structural changes. Herein, the structural effects of cysteine oxidation to the sulfinic acid were investigated in model peptides and folded proteins using NMR spectroscopy, circular dichroism, bioinformatics, and computational studies. In the PDB, Cys-SO2– shows a greater preference for α-helix than Cys. In addition, Cys-SO2– is more commonly found in structures with φ > 0, including in multiple types of β-turn. Sulfinate oxygens engage in hydrogen bonds with adjacent (i or i + 1) amide hydrogens. Over half of sulfinates have at least one hydrogen bond with an adjacent amide, and several structures have hydrogen bonds with both adjacent amides. Alternately, sulfur or either oxygen can act as an electron donor for n→π* interactions with the backbone carbonyl of the same residue, as indicated by frequent S⋯CO or O⋯CO distances below the sums of their van der Waals radii in protein structures. In peptides, Cys-SO2– favored α-helical structure at the N-terminus, consistent with helix dipole effects and backbone hydrogen bonds with the sulfinate promoting α-helix. Cys-SO2– has only modestly greater polyproline II helix propensity than Cys-SH, likely due to competition from multiple side chain-backbone interactions. Cys-SO2– stabilizes the i+1 position of a β-turn relative to Cys-SH. Within proteins, the range of side chain-main chain interactions available to Cys-SO2– compared to Cys-SH provides a basis for potential changes in protein structure and function due to cysteine oxidation to the sulfinic acid.

Protective Effect of Dietary Taurine from ROS Production in European Seabass under Conditions of Forced Swimming.

Simple SummaryA number of recent studies have demonstrated the essentiality of dietary taurine in many commercially relevant cultured fish species. Taurine is involved in many physiological functions in fish and represents an essential nutrient, which exert powerful antioxidant properties and is required as a supplement in the feed when a relevant percentage of vegetable protein sources are utilized. Our results show that dietary taurine can reduce the oxidative status of marine fish under swim stamina stress conditions. These data could be essential for the development of diets with fishmeal/soybean meal substitutions in the effort to improve the sustainability of the aquaculture.Taurine (Tau) is an amino sulfonic acid, which is widely distributed in animal tissues, whereas it is almost lacking in plants with the exception of certain algae, seaweeds, and few others. In the aquafeed industry, Tau is mainly used as a feed additive to promote growth in marine fish species with limited cysteine sulfinate decarboxylase activity. In particular, Tau supplementation is required in feeds in which fishmeal (FM) is substituted with high percentages of plant-derived protein sources such as soybean meals (SBM) that have much lower levels of Tau than FM. In addition to being a growth promoter, Tau exert powerful antioxidant properties being a scavenger of the reactive oxygen species (ROS). Under sustained swimming conditions, an intracellular increase in ROS production can occur in fish red muscle where the abundance of mitochondria (the main site of ROS formation) is high. Accordingly, this study aimed at investigating the effects of dietary Tau on European seabass (Dicentrarchus labrax) growth and oxidative stress response induced by swimming exercise. Individually tagged fish of 92.57 ± 20.33 g mean initial weight were fed two experimental diets containing the same low percentage of FM and high percentage of SBM. One diet was supplemented with 1.5% of Tau. Tau supplemented in the diet had a positive effect on fish growth, and enhanced swimming performance and antioxidant status. Two swim endurance tests were performed during the feeding trial. Metabolic oxygen consumption (MO2) was measured during exercise at incremental swimming speeds (0.7, 1.4, 2.1, 2.8, 3.5, and then 4.2 BL (body length) s−1, until fatigue). Fish maximal sustainable swimming speed (Ucrit) was determined too. To investigate the antioxidant effect of dietary Tau, we also measured ROS production in fish blood by RBA (respiratory burst activity) assay and quantified the expression of genes coding for antioxidant enzymes by qPCR (quantitative polymerase chain reaction) , such as SOD (superoxide dismutase), GPX (glutathione peroxidase), and CAT (catalase) in red muscle and liver. There was a significant effect of Tau upon Ucrit during exercise. Additionally, ROS production was significantly lower in fish fed with Tau supplemented diet, supporting the role of Tau as ROS scavenger. The protective effect of Tau against oxidative stress induced by forced swimming was denoted also by a significant decrease in antioxidant enzymes gene expression in fish liver and muscle. Taken together these results demonstrate that Tau is beneficial in low FM-based diets for seabass.

Uncoupling proteins 1 and 2 (UCP1 and UCP2) from Arabidopsis thaliana are mitochondrial transporters of aspartate, glutamate, and dicarboxylates

The Arabidopsis thaliana genome contains 58 members of the solute carrier family SLC25, also called the mitochondrial carrier family, many of which have been shown to transport specific metabolites, nucleotides, and cofactors across the mitochondrial membrane. Here, two Arabidopsis members of this family, AtUCP1 and AtUCP2, which were previously thought to be uncoupling proteins and hence named UCP1/PUMP1 and UCP2/PUMP2, respectively, are assigned with a novel function. They were expressed in bacteria, purified, and reconstituted in phospholipid vesicles. Their transport properties demonstrate that they transport amino acids (aspartate, glutamate, cysteine sulfinate, and cysteate), dicarboxylates (malate, oxaloacetate, and 2-oxoglutarate), phosphate, sulfate, and thiosulfate. Transport was saturable and inhibited by mercurials and other mitochondrial carrier inhibitors to various degrees. AtUCP1 and AtUCP2 catalyzed a fast counterexchange transport as well as a low uniport of substrates, with transport rates of AtUCP1 being much higher than those of AtUCP2 in both cases. The aspartate/glutamate heteroexchange mediated by AtUCP1 and AtUCP2 is electroneutral, in contrast to that mediated by the mammalian mitochondrial aspartate glutamate carrier. Furthermore, both carriers were found to be targeted to mitochondria. Metabolite profiling of single and double knockouts shows changes in organic acid and amino acid levels. Notably, AtUCP1 and AtUCP2 are the first reported mitochondrial carriers in Arabidopsis to transport aspartate and glutamate. It is proposed that the primary function of AtUCP1 and AtUCP2 is to catalyze an aspartateout/glutamatein exchange across the mitochondrial membrane and thereby contribute to the export of reducing equivalents from the mitochondria in photorespiration.

Identification and expression of cysteine sulfinate decarboxylase, possible regulation of taurine biosynthesis in Crassostrea gigas in response to low salinity

Taurine has been reported high amounts in marine animals to maintain osmotic balance between osmoformers and sea water. Approximately 80% of the total amino-acid content is taurine in Pacific oyster Crassostrea gigas, an intertidal and euryhaline species. In this study, we cloned the two copies of cysteine sulfinate decarboxylase (CSAD), the key enzyme in taurine biosynthesis pathway, screened in oyster genome data. Sequentially, we compared the expression patterns of CgCSAD1 and CgCSAD2 under low salinity treatment (8‰ and 15‰) using different families from two populations. There was no correlation between the expression of CSAD and the different population. Notably, CgCSAD1 increased significantly in treated groups for 24 h, but CgCSAD2 had no significant differentiation. Moreover, the results of CgCSAD1 interference provided the evidence of the positive correlation between CgCSAD1 expressions and taurine contents. The zinc finger domain showed in multi-alignment results may be the important character of CgCSAD1 as the key enzyme in taurine biosynthesis to regulate taurine pool in response to low salinity. This study provides a new evidence for the important role of taurine in adaptation to low salinity in oyster. In addition, it is a good model to discuss the function and evolution of the duplication in mollusks.

Taurine Biosynthesis in a Fish Liver Cell Line (ZFL) Adapted to a Serum-Free Medium.

Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity.

Differential regulation of taurine biosynthesis in rainbow trout and Japanese flounder.

Animals have varied taurine biosynthesis capability, which was determined by activities of key enzymes including cysteine dioxygenase (CDO) and cysteine sulfinate decarboxylase (CSD). However, whether CDO and CSD are differentially regulated across species remains unexplored. In the present study, we examined the regulations of CDO and CSD in rainbow trout and Japanese flounder, the two fish species with high and low taurine biosynthesis ability respectively. Our results showed that the expression of CDO was lower in rainbow trout but more responsive to cysteine stimulation compared to that in Japanese flounder. On the other hand, both the expression and catalytic efficiency (kcat) of CSD were higher in rainbow trout than those of Japanese flounder. A three-residue substrate recognition motif in rainbow trout CSD with sequence of F126/S146/Y148 was identified to be responsible for high kcat, while that with sequence of F88/N108/F110 in Japanese flounder led to low kcat, as suggested by site-directed mutagenesis studies. In summary, our results determined new aspects of taurine biosynthesis regulation across species.

Radical Formation in the Gas-Phase Ozonolysis of Deprotonated Cysteine

Although the deleterious effects of ozone on the human respiratory system are well-known, many of the precise chemical mechanisms that both cause damage and afford protection in the pulmonary epithelial lining fluid are poorly understood. As a key first step to elucidating the intrinsic reactivity of ozone with proteins, its reactions with deprotonated cysteine [Cys-H]- are examined in the gas phase. Reaction proceeds at near the collision limit to give a rich set of products including 1) sequential oxygen atom abstraction reactions to yield cysteine sulfenate, sulfinate and sulfonate anions, and significantly 2) sulfenate radical anions formed by ejection of a hydroperoxy radical. The free-radical pathway occurs only when both thiol and carboxylate moieties are available, implicating electron-transfer as a key step in this reaction. This novel and facile reaction is also observed in small cys-containing peptides indicating a possible role for this chemistry in protein ozonolysis. Gas-phase reactions: Deprotonated cysteine reacted with ozone to give a rich set of products including sequential oxygen atom abstraction reactions to yield cysteine sulfenate, sulfonate, and sulfonate anions, and significantly sulfenate radical anions formed by ejection of a hydroperoxy radical (see picture). The free-radical pathway occurred only when both thiol and carboxylate moieties were available.

Radical Formation in the Gas-Phase Ozonolysis of Deprotonated Cysteine.

Although the deleterious effects of ozone on the human respiratory system are well-known, many of the precise chemical mechanisms that both cause damage and afford protection in the pulmonary epithelial lining fluid are poorly understood. As a key first step to elucidating the intrinsic reactivity of ozone with proteins, its reactions with deprotonated cysteine [Cys-H](-) are examined in the gas phase. Reaction proceeds at near the collision limit to give a rich set of products including 1) sequential oxygen atom abstraction reactions to yield cysteine sulfenate, sulfinate and sulfonate anions, and significantly 2) sulfenate radical anions formed by ejection of a hydroperoxy radical. The free-radical pathway occurs only when both thiol and carboxylate moieties are available, implicating electron-transfer as a key step in this reaction. This novel and facile reaction is also observed in small cys-containing peptides indicating a possible role for this chemistry in protein ozonolysis.

Taurine deficiency, synthesis and transport in the mdx mouse model for Duchenne Muscular Dystrophy

The amino acid taurine is essential for the function of skeletal muscle and administration is proposed as a treatment for Duchenne Muscular Dystrophy (DMD). Taurine homeostasis is dependent on multiple processes including absorption of taurine from food, endogenous synthesis from cysteine and reabsorption in the kidney. This study investigates the cause of reported taurine deficiency in the dystrophic mdx mouse model of DMD. Levels of metabolites (taurine, cysteine, cysteine sulfinate and hypotaurine) and proteins (taurine transporter [TauT], cysteine deoxygenase and cysteine sulfinate dehydrogenase) were quantified in juvenile control C57 and dystrophic mdx mice aged 18 days, 4 and 6 weeks. In C57 mice, taurine content was much higher in both liver and plasma at 18 days, and both cysteine and cysteine deoxygenase were increased. As taurine levels decreased in maturing C57 mice, there was increased transport (reabsorption) of taurine in the kidney and muscle. In mdx mice, taurine and cysteine levels were much lower in liver and plasma at 18 days, and in muscle cysteine was low at 18 days, whereas taurine was lower at 4: these changes were associated with perturbations in taurine transport in liver, kidney and muscle and altered metabolism in liver and kidney. These data suggest that the maintenance of adequate body taurine relies on sufficient dietary intake of taurine and cysteine availability and metabolism, as well as retention of taurine by the kidney. This research indicates dystrophin deficiency not only perturbs taurine metabolism in the muscle but also affects taurine metabolism in the liver and kidney, and supports targeting cysteine and taurine deficiency as a potential therapy for DMD.