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Robert D. Steele - One of the best experts on this subject based on the ideXlab platform.
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Cloning and characterization of rat Cysteine Sulfinic Acid decarboxylase
Biochimica et biophysica acta, 1995Co-Authors: Pamela J. Kaisakia, Ann A. Jerkins, David C. Goodspeed, Robert D. SteeleAbstract:Cysteine Sulfinic Acid decarboxylase (CSAD) is a key enzyme in taurine biosynthesis. CSAD activity and enzyme protein concentration are both repressed by the action of the steroid family hormones triiodothyronine and estrogen. To characterize this suppression, a cDNA clone for CSAD was isolated from a rat liver cDNA expression library using polyclonal antibodies to CSAD. The cDNA was sequenced in its entirety and confirmed to be a clone of CSAD. In a Northern blot comparing liver and kidney RNA of male and female rats, the CSAD cDNA probe detected a 2.5 kb mRNA band which was present at levels corresponding to the concentration of enzyme protein. Hyperthyroidism decreased CSAD mRNA as compared to euthyroid controls, providing evidence that negative regulation of CSAD activity occurs at the level of mRNA.
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quantification of Cysteine Sulfinic Acid decar ylase in male and female rats effect of adrenalectomy and methionine
Archives of Biochemistry and Biophysics, 1992Co-Authors: Ann A. Jerkins, Robert D. SteeleAbstract:Abstract Hepatic Cysteine Sulfinic Acid decar☐ylase (EC 4.1.1.29) activity has been reported to decrease in response to both l -methionine (Met) feeding and adrenalectomy in rats. A series of experiments was conducted to (a) determine if CSAD depression was evident in female rats fed a methionine-supplemented diet; and (b) determine if adrenal hormones mediated the response of CSAD to dietary methionine. Cysteine Sulfinic Acid decar☐ylase (CSAD) activity was measured in livers of male and female rats fed a methionine-supplemented diet. In female rat liver, CSAD activity was only 25% of the activity measured in livers of male rats. Hepatic enzyme activity in male rats fed a casein-based basal diet containing 0.6% l -methionine was 2.5-fold higher than activity in male rats fed a methionine-supplemented diet containing 1.35% l -methionine (+Met). Similarly, enzyme activity in livers of female rats fed the basal diet was 1.7-fold higher than in female rats fed a methionine-supplemented diet. CSAD activity in adrenalectomized (ADX) male rats fed the basal diet was depressed (990 ± 120 nmol/min · g liver) compared to activity in intact controls (2347 ± 89) and sham controls (2040 ± 143) fed the basal diet. CSAD activity was further depressed in ADX, intact controls, and sham controls fed +Met. Immunochemical detection and quantification of CSAD protein in rat liver demonstrated that changes in CSAD protein were consistent with the observed decreased enzyme activity in female rats, ADX rats, and rats fed +Met. S -Adenosylmethionine and S -adenosylhomoCysteine concentrations tended to increase in livers of rats fed +Met. ADX rats fed +Met had the greatest increase in S -adenosylmethionine and S -adenosylhomoCysteine concentrations. The depression in hepatic CSAD observed after feeding +Met to rats does not appear to involve adrenal function.
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Quantification of Cysteine Sulfinic Acid decar☐ylase in male and female rats: Effect of adrenalectomy and methionine
Archives of biochemistry and biophysics, 1992Co-Authors: Ann A. Jerkins, Robert D. SteeleAbstract:Abstract Hepatic Cysteine Sulfinic Acid decar☐ylase (EC 4.1.1.29) activity has been reported to decrease in response to both l -methionine (Met) feeding and adrenalectomy in rats. A series of experiments was conducted to (a) determine if CSAD depression was evident in female rats fed a methionine-supplemented diet; and (b) determine if adrenal hormones mediated the response of CSAD to dietary methionine. Cysteine Sulfinic Acid decar☐ylase (CSAD) activity was measured in livers of male and female rats fed a methionine-supplemented diet. In female rat liver, CSAD activity was only 25% of the activity measured in livers of male rats. Hepatic enzyme activity in male rats fed a casein-based basal diet containing 0.6% l -methionine was 2.5-fold higher than activity in male rats fed a methionine-supplemented diet containing 1.35% l -methionine (+Met). Similarly, enzyme activity in livers of female rats fed the basal diet was 1.7-fold higher than in female rats fed a methionine-supplemented diet. CSAD activity in adrenalectomized (ADX) male rats fed the basal diet was depressed (990 ± 120 nmol/min · g liver) compared to activity in intact controls (2347 ± 89) and sham controls (2040 ± 143) fed the basal diet. CSAD activity was further depressed in ADX, intact controls, and sham controls fed +Met. Immunochemical detection and quantification of CSAD protein in rat liver demonstrated that changes in CSAD protein were consistent with the observed decreased enzyme activity in female rats, ADX rats, and rats fed +Met. S -Adenosylmethionine and S -adenosylhomoCysteine concentrations tended to increase in livers of rats fed +Met. ADX rats fed +Met had the greatest increase in S -adenosylmethionine and S -adenosylhomoCysteine concentrations. The depression in hepatic CSAD observed after feeding +Met to rats does not appear to involve adrenal function.
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Cysteine Sulfinic Acid decarboxylase activity in response to thyroid hormone administration in rats
Archives of biochemistry and biophysics, 1991Co-Authors: Ann A. Jerkins, Robert D. SteeleAbstract:The modulation of hepatic and renal Cysteine Sulfinic Acid decarboxylase (EC 4.1.1.29) activities by triiodothyronine (T3) was studied in a series of experiments. In a dose--response study, hepatic Cysteine Sulfinic Acid decarboxylase activity (CSAD) was depressed by 65% and renal activity was increased threefold in rats injected with 100 micrograms T3/100 g body wt for 7 days when compared to rats injected with 0.3 micrograms T3/100 g body wt. Western blot analysis indicated that these changes in CSAD activity were due to changes in the quantity of CSAD protein. Changes in hepatic and renal activities were not evident until 24 h after T3 administration. In response to T3 clearance, hepatic and renal CSAD activities approached euthyroid values 4-7 days after cessation of T3 injections although serum T3 concentrations were no different from euthyroid values 48 h after T3 injections were stopped. These data indicate that thyroid hormone effects persist after T3 clearance. The response of CSAD to thyroid status may be related to its role in taurine biosynthesis.
Georgia Schuller-levis - One of the best experts on this subject based on the ideXlab platform.
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A Novel Cysteine Sulfinic Acid Decarboxylase Knock-Out Mouse: Immune Function (II).
Advances in experimental medicine and biology, 2017Co-Authors: Eunkyue Park, Seung Yong Park, In Soo Cho, Bo Sook Kim, Georgia Schuller-levisAbstract:Taurine deficient mice lacking Cysteine Sulfinic Acid decarboxylase (CSAD KO) were developed for investigating the various physiological roles of taurine including the development of the brain and eye as well as immune function. Due to severe abnormalities of immune function in a taurine deficient cat, the immune function including adoptive and innate immunity in taurine-deficient mice have been studied. Previously we demonstrated that B cell function in CSAD KO was reduced in both females and males. However, T cell function was significantly reduced only in females. In this study, we have examined innate immunity using macrophage activation with LPS or/and IFN-γ and polymorphonuclear leukocytes (PMN) activation with phorbol myristate acetate (PMA). Pro- and anti-inflammatory cytokines including IL-6, TNF-α and IL-10 as well as nitric oxide (NO) were determined using ELISA and Griess reagent, respectively. Peritoneal macrophages were activated with 1 μg/mL of lipopolysaccharide (LPS) and/or 50 U/mL of IFN-γ. In addition, superoxide anion was measured using peritoneal PMN activated with PMA in the presence and absence of superoxide dismutase. Superoxide anion production in activated PMN from CSAD KO homozygotes (HO) was not significantly different from wild-type (WT) with and without 25 mM taurine. IL-10 and TNF-α production in both female and male CSAD KO were not significantly different. IL-6 and NO were significantly lower only in females as previously observed in Con A-activated cellular proliferation of splenocytes. Cytokine production with 10 mM of taurine was not different, indicating the reduction of NO and IL-6 in females may be due to the absence of the CSAD gene, not due to low taurine concentrations.These data indicate that some measures of innate immunity were altered in female CSAD mice.
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A Novel Cysteine Sulfinic Acid Decarboxylase Knock-Out Mouse: Taurine Distribution in Various Tissues With and Without Taurine Supplementation
Advances in experimental medicine and biology, 2017Co-Authors: Eunkyue Park, Seung Yong Park, In Soo Cho, Bo Sook Kim, Georgia Schuller-levisAbstract:Taurine, a sulfur containing amino Acid, has various physiological functions including development of the eye and brain, immune function, reproduction, osmo-regulatory function as well as anti-oxidant and anti-inflammatory activities. In order to understand the physiological role, we developed taurine deficient mice deleting a rate-liming enzyme, Cysteine Sulfinic Acid decarboxylase (CSAD) for biosynthesis of taurine. Taurine was measured in various tissues including the liver, brain, lung, spleen, thymus, pancreas, heart, muscle and kidney as well as plasma from CSAD knock-out mice (CSAD KO) with and without treatment of taurine in the drinking water at the age of 2 months (2 M). Taurine was determined using HPLC as a phenylisothiocyanate derivative of taurine at 254 nm. Taurine concentrations in the liver and kidney from homozygotes of CSAD KO (HO), in which CSAD level is high, were 90% and 70% lower than WT, respectively. Taurine concentrations in the brain, spleen and lung, where CSAD level is low, were 21%, 20% and 28% lower than WT, respectively. At 2 M, 1% taurine treatment of HO restored taurine concentrations in all tissues compared to that of WT. To select an appropriate taurine treatment, HO were treated with various concentrations (0.05, 0.2, 1%) of taurine for 4 months (4 M). Restoration of taurine in all tissues except the liver, kidney and lung requires 0.05% taurine to be restored to that of WT. The liver and kidney restore taurine back to WT with 0.2% taurine. To examine which enzymes influence taurine concentrations in various tissues from WT and HO at 2 M, expression of five taurine-related enzymes, two antioxidant enzymes as well as lactoferrin (Lft) and prolactin receptor (Prlr) was determined using RT2 qPCR. The expression of taurine transporter in the liver, brain, muscle and kidney from HO was increased except in the lung. Our data showed expression of glutamate decarboxylase-like 1(Gadl-1) was increased in the brain and muscle in HO, compared to WT, indicating taurine in the brain and muscle from HO was replenished through taurine transporter and increased biosynthesis of taurine by up-regulated Gadl-1. The expression of glutathione peroxidase 3 was increased in the brain and peroxireductase 2 was increased in the liver and lung, suggesting taurine has anti-oxidant activity. In contrast to newborn and 1 month CSAD KO, Ltf and Prlr in the liver from CSAD KO at 2 M were increased more than two times and 52%, respectively, indicating these two proteins may be required for pregnancy of CSAD KO. Ltf in HOT1.0 was restored to WT, while Prlr in HOT1.0 was increased more than HO, explaining improvement of neonatal survival with taurine supplementation.
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A novel Cysteine Sulfinic Acid decarboxylase knock-out mouse: immune function.
Advances in experimental medicine and biology, 2015Co-Authors: Seung Yong Park, Georgia Schuller-levis, Eunkyue ParkAbstract:Taurine deficient mice lacking Cysteine Sulfinic Acid decarboxylase (CSAD KO) were developed for investigating the various physiological roles of taurine including the development of the brain and eye as well as immune function. Due to severe abnormalities of immune function in a taurine deficient cat, the immune function including adoptive and innate immunity in taurine-deficient mice have been studied. Previously we demonstrated that B cell function in CSAD KO was reduced in both females and males. However, T cell function was significantly reduced only in females. In this study, we have examined innate immunity using macrophage activation with LPS or/and IFN-γ and polymorphonuclear leukocytes (PMN) activation with phorbol myristate acetate (PMA). Pro- and anti-inflammatory cytokines including IL-6, TNF-α and IL-10 as well as nitric oxide (NO) were determined using ELISA and Griess reagent, respectively. Peritoneal macrophages were activated with 1 μg/mL of lipopolysaccharide (LPS) and/or 50 U/mL of IFN-γ. In addition, superoxide anion was measured using peritoneal PMN activated with PMA in the presence and absence of superoxide dismutase. Superoxide anion production in activated PMN from CSAD KO homozygotes (HO) was not significantly different from wild-type (WT) with and without 25 mM taurine. IL-10 and TNF-α production in both female and male CSAD KO were not significantly different. IL-6 and NO were significantly lower only in females as previously observed in Con A-activated cellular proliferation of splenocytes. Cytokine production with 10 mM of taurine was not different, indicating the reduction of NO and IL-6 in females may be due to the absence of the CSAD gene, not due to low taurine concentrations.
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A novel Cysteine Sulfinic Acid decarboxylase knock-out mouse: comparison between newborn and weanling mice.
Advances in experimental medicine and biology, 2015Co-Authors: Eunkyue Park, Seung Yong Park, Carl Dobkin, Georgia Schuller-levisAbstract:We developed a novel Cysteine Sulfinic Acid decarboxylase knockout mouse (CSAD KO) to investigate the critical roles of taurine. The absence of the CSAD gene was confirmed using Southern, northern and western blotting; homozygous (CSAD−/−) and heterozygous (CSAD+/-) animals were identified using PCR. Plasma taurine concentrations were decreased by 86 % in CSAD−/− animals. Reproductive performance was poor in the second and later CSAD−/− generations but was restored by supplementing the drinking water with 0.05 % taurine. Taurine concentrations at postnatal day 1 (PD1) in generation 1 (G1) CSAD−/− were close to normal presumably due to taurine transport through the placenta from the CSAD+/− dam. In contrast, taurine concentrations in the brain and liver of G2, G3, and G4 CSAD−/− were very low at birth. At 1 month of age (1 M), 1 week after weaning, the G1 CSAD−/− taurine concentration in the liver decreased to the same level as G2, G3, and G4 CSAD−/−. Taurine concentrations in all CSAD−/− at 1 M were reduced to 44 % of WT in the brain and 5 % in the liver. Taurine supplementation restored brain levels to 76 % of WT but had little effect on the liver. Gene expression in CSAD−/− brain and liver was compared to WT at PD1 and 1 M. Expression of the prolactin receptor and lactoferrin genes was decreased in CSAD−/− at both PD1 and 1 M. Metabolic genes including serine dehydratase and uridine phosphorylase 2 were increased significantly at PD1 but not at 1 M. An oxidative stress gene, glutathioneperoxidase 3 was increased in CSAD KO−/− at both PD1 and 1 M. Expression of taurine metabolism genes, Cysteine dioxygenase (Cdo), cysteamine dioxygenase and the taurine transporter (TauT) were unaffected at PD1. At 1 M, however, TauT in CSAD−/− liver was increased twofold, while Cdo was decreased compared to WT. These data indicate that the CSAD KO mouse is a powerful model for exploring the role of taurine deficiency in various disorders especially since the requirement for taurine can be supplied in food or water.
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Cloning of murine Cysteine Sulfinic Acid decarboxylase and its mRNA expression in murine tissues
Biochimica et biophysica acta, 2002Co-Authors: Eunkyue Park, Seung Yong Park, Chuanhua Wang, Giuseppe Lafauci, Georgia Schuller-levisAbstract:Cysteine Sulfinic Acid decarboxylase (CSD) is the rate-limiting enzyme for biosynthesis of taurine which is essential to biological processes such as development of the brain and eye, reproduction, osmoregulation as well as the anti-inflammatory activity of leukocytes. We report the cDNA sequence of murine CSD that predicts a polypeptide of 493 amino Acids. This protein shares 98% and 90% of amino Acids with rat and human CSD, respectively, indicating that it is a true ortholog of CSD. Northern blot analysis revealed that CSD mRNA is expressed in kidney and liver, and was not detected in lymphoid tissues and lung. The nucleotide sequence of murine CSD should be useful for genetic manipulation of the CSD gene.
Eunkyue Park - One of the best experts on this subject based on the ideXlab platform.
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A Novel Cysteine Sulfinic Acid Decarboxylase Knock-Out Mouse: Immune Function (II).
Advances in experimental medicine and biology, 2017Co-Authors: Eunkyue Park, Seung Yong Park, In Soo Cho, Bo Sook Kim, Georgia Schuller-levisAbstract:Taurine deficient mice lacking Cysteine Sulfinic Acid decarboxylase (CSAD KO) were developed for investigating the various physiological roles of taurine including the development of the brain and eye as well as immune function. Due to severe abnormalities of immune function in a taurine deficient cat, the immune function including adoptive and innate immunity in taurine-deficient mice have been studied. Previously we demonstrated that B cell function in CSAD KO was reduced in both females and males. However, T cell function was significantly reduced only in females. In this study, we have examined innate immunity using macrophage activation with LPS or/and IFN-γ and polymorphonuclear leukocytes (PMN) activation with phorbol myristate acetate (PMA). Pro- and anti-inflammatory cytokines including IL-6, TNF-α and IL-10 as well as nitric oxide (NO) were determined using ELISA and Griess reagent, respectively. Peritoneal macrophages were activated with 1 μg/mL of lipopolysaccharide (LPS) and/or 50 U/mL of IFN-γ. In addition, superoxide anion was measured using peritoneal PMN activated with PMA in the presence and absence of superoxide dismutase. Superoxide anion production in activated PMN from CSAD KO homozygotes (HO) was not significantly different from wild-type (WT) with and without 25 mM taurine. IL-10 and TNF-α production in both female and male CSAD KO were not significantly different. IL-6 and NO were significantly lower only in females as previously observed in Con A-activated cellular proliferation of splenocytes. Cytokine production with 10 mM of taurine was not different, indicating the reduction of NO and IL-6 in females may be due to the absence of the CSAD gene, not due to low taurine concentrations.These data indicate that some measures of innate immunity were altered in female CSAD mice.
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A Novel Cysteine Sulfinic Acid Decarboxylase Knock-Out Mouse: Taurine Distribution in Various Tissues With and Without Taurine Supplementation
Advances in experimental medicine and biology, 2017Co-Authors: Eunkyue Park, Seung Yong Park, In Soo Cho, Bo Sook Kim, Georgia Schuller-levisAbstract:Taurine, a sulfur containing amino Acid, has various physiological functions including development of the eye and brain, immune function, reproduction, osmo-regulatory function as well as anti-oxidant and anti-inflammatory activities. In order to understand the physiological role, we developed taurine deficient mice deleting a rate-liming enzyme, Cysteine Sulfinic Acid decarboxylase (CSAD) for biosynthesis of taurine. Taurine was measured in various tissues including the liver, brain, lung, spleen, thymus, pancreas, heart, muscle and kidney as well as plasma from CSAD knock-out mice (CSAD KO) with and without treatment of taurine in the drinking water at the age of 2 months (2 M). Taurine was determined using HPLC as a phenylisothiocyanate derivative of taurine at 254 nm. Taurine concentrations in the liver and kidney from homozygotes of CSAD KO (HO), in which CSAD level is high, were 90% and 70% lower than WT, respectively. Taurine concentrations in the brain, spleen and lung, where CSAD level is low, were 21%, 20% and 28% lower than WT, respectively. At 2 M, 1% taurine treatment of HO restored taurine concentrations in all tissues compared to that of WT. To select an appropriate taurine treatment, HO were treated with various concentrations (0.05, 0.2, 1%) of taurine for 4 months (4 M). Restoration of taurine in all tissues except the liver, kidney and lung requires 0.05% taurine to be restored to that of WT. The liver and kidney restore taurine back to WT with 0.2% taurine. To examine which enzymes influence taurine concentrations in various tissues from WT and HO at 2 M, expression of five taurine-related enzymes, two antioxidant enzymes as well as lactoferrin (Lft) and prolactin receptor (Prlr) was determined using RT2 qPCR. The expression of taurine transporter in the liver, brain, muscle and kidney from HO was increased except in the lung. Our data showed expression of glutamate decarboxylase-like 1(Gadl-1) was increased in the brain and muscle in HO, compared to WT, indicating taurine in the brain and muscle from HO was replenished through taurine transporter and increased biosynthesis of taurine by up-regulated Gadl-1. The expression of glutathione peroxidase 3 was increased in the brain and peroxireductase 2 was increased in the liver and lung, suggesting taurine has anti-oxidant activity. In contrast to newborn and 1 month CSAD KO, Ltf and Prlr in the liver from CSAD KO at 2 M were increased more than two times and 52%, respectively, indicating these two proteins may be required for pregnancy of CSAD KO. Ltf in HOT1.0 was restored to WT, while Prlr in HOT1.0 was increased more than HO, explaining improvement of neonatal survival with taurine supplementation.
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A novel Cysteine Sulfinic Acid decarboxylase knock-out mouse: immune function.
Advances in experimental medicine and biology, 2015Co-Authors: Seung Yong Park, Georgia Schuller-levis, Eunkyue ParkAbstract:Taurine deficient mice lacking Cysteine Sulfinic Acid decarboxylase (CSAD KO) were developed for investigating the various physiological roles of taurine including the development of the brain and eye as well as immune function. Due to severe abnormalities of immune function in a taurine deficient cat, the immune function including adoptive and innate immunity in taurine-deficient mice have been studied. Previously we demonstrated that B cell function in CSAD KO was reduced in both females and males. However, T cell function was significantly reduced only in females. In this study, we have examined innate immunity using macrophage activation with LPS or/and IFN-γ and polymorphonuclear leukocytes (PMN) activation with phorbol myristate acetate (PMA). Pro- and anti-inflammatory cytokines including IL-6, TNF-α and IL-10 as well as nitric oxide (NO) were determined using ELISA and Griess reagent, respectively. Peritoneal macrophages were activated with 1 μg/mL of lipopolysaccharide (LPS) and/or 50 U/mL of IFN-γ. In addition, superoxide anion was measured using peritoneal PMN activated with PMA in the presence and absence of superoxide dismutase. Superoxide anion production in activated PMN from CSAD KO homozygotes (HO) was not significantly different from wild-type (WT) with and without 25 mM taurine. IL-10 and TNF-α production in both female and male CSAD KO were not significantly different. IL-6 and NO were significantly lower only in females as previously observed in Con A-activated cellular proliferation of splenocytes. Cytokine production with 10 mM of taurine was not different, indicating the reduction of NO and IL-6 in females may be due to the absence of the CSAD gene, not due to low taurine concentrations.
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A novel Cysteine Sulfinic Acid decarboxylase knock-out mouse: comparison between newborn and weanling mice.
Advances in experimental medicine and biology, 2015Co-Authors: Eunkyue Park, Seung Yong Park, Carl Dobkin, Georgia Schuller-levisAbstract:We developed a novel Cysteine Sulfinic Acid decarboxylase knockout mouse (CSAD KO) to investigate the critical roles of taurine. The absence of the CSAD gene was confirmed using Southern, northern and western blotting; homozygous (CSAD−/−) and heterozygous (CSAD+/-) animals were identified using PCR. Plasma taurine concentrations were decreased by 86 % in CSAD−/− animals. Reproductive performance was poor in the second and later CSAD−/− generations but was restored by supplementing the drinking water with 0.05 % taurine. Taurine concentrations at postnatal day 1 (PD1) in generation 1 (G1) CSAD−/− were close to normal presumably due to taurine transport through the placenta from the CSAD+/− dam. In contrast, taurine concentrations in the brain and liver of G2, G3, and G4 CSAD−/− were very low at birth. At 1 month of age (1 M), 1 week after weaning, the G1 CSAD−/− taurine concentration in the liver decreased to the same level as G2, G3, and G4 CSAD−/−. Taurine concentrations in all CSAD−/− at 1 M were reduced to 44 % of WT in the brain and 5 % in the liver. Taurine supplementation restored brain levels to 76 % of WT but had little effect on the liver. Gene expression in CSAD−/− brain and liver was compared to WT at PD1 and 1 M. Expression of the prolactin receptor and lactoferrin genes was decreased in CSAD−/− at both PD1 and 1 M. Metabolic genes including serine dehydratase and uridine phosphorylase 2 were increased significantly at PD1 but not at 1 M. An oxidative stress gene, glutathioneperoxidase 3 was increased in CSAD KO−/− at both PD1 and 1 M. Expression of taurine metabolism genes, Cysteine dioxygenase (Cdo), cysteamine dioxygenase and the taurine transporter (TauT) were unaffected at PD1. At 1 M, however, TauT in CSAD−/− liver was increased twofold, while Cdo was decreased compared to WT. These data indicate that the CSAD KO mouse is a powerful model for exploring the role of taurine deficiency in various disorders especially since the requirement for taurine can be supplied in food or water.
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Cloning of murine Cysteine Sulfinic Acid decarboxylase and its mRNA expression in murine tissues
Biochimica et biophysica acta, 2002Co-Authors: Eunkyue Park, Seung Yong Park, Chuanhua Wang, Giuseppe Lafauci, Georgia Schuller-levisAbstract:Cysteine Sulfinic Acid decarboxylase (CSD) is the rate-limiting enzyme for biosynthesis of taurine which is essential to biological processes such as development of the brain and eye, reproduction, osmoregulation as well as the anti-inflammatory activity of leukocytes. We report the cDNA sequence of murine CSD that predicts a polypeptide of 493 amino Acids. This protein shares 98% and 90% of amino Acids with rat and human CSD, respectively, indicating that it is a true ortholog of CSD. Northern blot analysis revealed that CSD mRNA is expressed in kidney and liver, and was not detected in lymphoid tissues and lung. The nucleotide sequence of murine CSD should be useful for genetic manipulation of the CSD gene.
Ann A. Jerkins - One of the best experts on this subject based on the ideXlab platform.
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Cysteine Sulfinic Acid Decarboxylase mRNA Abundance Decreases in Rats Fed a High-Protein Diet
The Journal of nutrition, 1998Co-Authors: Ann A. Jerkins, Deborah D. Jones, Edwin A. KohlheppAbstract:The partitioning of Cysteine metabolism between sulfate and taurine biosynthetic pathways may be regulated in part by the activity of Cysteine Sulfinic Acid decarboxylase (CSAD). CSAD activity is repressed by high-protein feeding, and we have previously reported that changes in CSAD activity are correlated with changes in CSAD protein. We conducted experiments to determine the relative expression of CSAD mRNA in rats fed 18 or 60% casein diets. In rats fed a 60% casein diet for 1 wk, hepatic CSAD activity and CSAD protein were 16 and 36%, respectively, of the values measured in rats fed the 18% casein diet. CSAD mRNA abundance in rats fed the 60% casein diet was 14% of the CSAD mRNA abundance in rats fed an 18% casein diet. The time course of the change in CSAD activity and mRNA abundance was examined in rats fed 18 or 60% casein diets for 48 h. Within 6 h of switching rats to a 60% casein diet, CSAD activity was decreased by 20% and after 48 h, activity was decreased 47% compared to activity measured at baseline. CSAD mRNA abundance was decreased 54% within 12 h of feeding rats a high-protein diet and remained depressed at 48 h. In a parallel group of rats fed the 18% casein diet, CSAD activity and CSAD mRNA were not significantly different from baseline values at 48 h. The decreased expression of CSAD mRNA in rats fed a high-protein diet is consistent with decreases in both CSAD enzyme activity and CSAD protein. Our results suggest dietary protein may regulate CSAD at the level of mRNA.
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Cloning and characterization of rat Cysteine Sulfinic Acid decarboxylase
Biochimica et biophysica acta, 1995Co-Authors: Pamela J. Kaisakia, Ann A. Jerkins, David C. Goodspeed, Robert D. SteeleAbstract:Cysteine Sulfinic Acid decarboxylase (CSAD) is a key enzyme in taurine biosynthesis. CSAD activity and enzyme protein concentration are both repressed by the action of the steroid family hormones triiodothyronine and estrogen. To characterize this suppression, a cDNA clone for CSAD was isolated from a rat liver cDNA expression library using polyclonal antibodies to CSAD. The cDNA was sequenced in its entirety and confirmed to be a clone of CSAD. In a Northern blot comparing liver and kidney RNA of male and female rats, the CSAD cDNA probe detected a 2.5 kb mRNA band which was present at levels corresponding to the concentration of enzyme protein. Hyperthyroidism decreased CSAD mRNA as compared to euthyroid controls, providing evidence that negative regulation of CSAD activity occurs at the level of mRNA.
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quantification of Cysteine Sulfinic Acid decar ylase in male and female rats effect of adrenalectomy and methionine
Archives of Biochemistry and Biophysics, 1992Co-Authors: Ann A. Jerkins, Robert D. SteeleAbstract:Abstract Hepatic Cysteine Sulfinic Acid decar☐ylase (EC 4.1.1.29) activity has been reported to decrease in response to both l -methionine (Met) feeding and adrenalectomy in rats. A series of experiments was conducted to (a) determine if CSAD depression was evident in female rats fed a methionine-supplemented diet; and (b) determine if adrenal hormones mediated the response of CSAD to dietary methionine. Cysteine Sulfinic Acid decar☐ylase (CSAD) activity was measured in livers of male and female rats fed a methionine-supplemented diet. In female rat liver, CSAD activity was only 25% of the activity measured in livers of male rats. Hepatic enzyme activity in male rats fed a casein-based basal diet containing 0.6% l -methionine was 2.5-fold higher than activity in male rats fed a methionine-supplemented diet containing 1.35% l -methionine (+Met). Similarly, enzyme activity in livers of female rats fed the basal diet was 1.7-fold higher than in female rats fed a methionine-supplemented diet. CSAD activity in adrenalectomized (ADX) male rats fed the basal diet was depressed (990 ± 120 nmol/min · g liver) compared to activity in intact controls (2347 ± 89) and sham controls (2040 ± 143) fed the basal diet. CSAD activity was further depressed in ADX, intact controls, and sham controls fed +Met. Immunochemical detection and quantification of CSAD protein in rat liver demonstrated that changes in CSAD protein were consistent with the observed decreased enzyme activity in female rats, ADX rats, and rats fed +Met. S -Adenosylmethionine and S -adenosylhomoCysteine concentrations tended to increase in livers of rats fed +Met. ADX rats fed +Met had the greatest increase in S -adenosylmethionine and S -adenosylhomoCysteine concentrations. The depression in hepatic CSAD observed after feeding +Met to rats does not appear to involve adrenal function.
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Quantification of Cysteine Sulfinic Acid decar☐ylase in male and female rats: Effect of adrenalectomy and methionine
Archives of biochemistry and biophysics, 1992Co-Authors: Ann A. Jerkins, Robert D. SteeleAbstract:Abstract Hepatic Cysteine Sulfinic Acid decar☐ylase (EC 4.1.1.29) activity has been reported to decrease in response to both l -methionine (Met) feeding and adrenalectomy in rats. A series of experiments was conducted to (a) determine if CSAD depression was evident in female rats fed a methionine-supplemented diet; and (b) determine if adrenal hormones mediated the response of CSAD to dietary methionine. Cysteine Sulfinic Acid decar☐ylase (CSAD) activity was measured in livers of male and female rats fed a methionine-supplemented diet. In female rat liver, CSAD activity was only 25% of the activity measured in livers of male rats. Hepatic enzyme activity in male rats fed a casein-based basal diet containing 0.6% l -methionine was 2.5-fold higher than activity in male rats fed a methionine-supplemented diet containing 1.35% l -methionine (+Met). Similarly, enzyme activity in livers of female rats fed the basal diet was 1.7-fold higher than in female rats fed a methionine-supplemented diet. CSAD activity in adrenalectomized (ADX) male rats fed the basal diet was depressed (990 ± 120 nmol/min · g liver) compared to activity in intact controls (2347 ± 89) and sham controls (2040 ± 143) fed the basal diet. CSAD activity was further depressed in ADX, intact controls, and sham controls fed +Met. Immunochemical detection and quantification of CSAD protein in rat liver demonstrated that changes in CSAD protein were consistent with the observed decreased enzyme activity in female rats, ADX rats, and rats fed +Met. S -Adenosylmethionine and S -adenosylhomoCysteine concentrations tended to increase in livers of rats fed +Met. ADX rats fed +Met had the greatest increase in S -adenosylmethionine and S -adenosylhomoCysteine concentrations. The depression in hepatic CSAD observed after feeding +Met to rats does not appear to involve adrenal function.
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Cysteine Sulfinic Acid decarboxylase activity in response to thyroid hormone administration in rats
Archives of biochemistry and biophysics, 1991Co-Authors: Ann A. Jerkins, Robert D. SteeleAbstract:The modulation of hepatic and renal Cysteine Sulfinic Acid decarboxylase (EC 4.1.1.29) activities by triiodothyronine (T3) was studied in a series of experiments. In a dose--response study, hepatic Cysteine Sulfinic Acid decarboxylase activity (CSAD) was depressed by 65% and renal activity was increased threefold in rats injected with 100 micrograms T3/100 g body wt for 7 days when compared to rats injected with 0.3 micrograms T3/100 g body wt. Western blot analysis indicated that these changes in CSAD activity were due to changes in the quantity of CSAD protein. Changes in hepatic and renal activities were not evident until 24 h after T3 administration. In response to T3 clearance, hepatic and renal CSAD activities approached euthyroid values 4-7 days after cessation of T3 injections although serum T3 concentrations were no different from euthyroid values 48 h after T3 injections were stopped. These data indicate that thyroid hormone effects persist after T3 clearance. The response of CSAD to thyroid status may be related to its role in taurine biosynthesis.
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Sestrin 2 is not a reductase for Cysteine Sulfinic Acid of peroxiredoxins.
Antioxidants & redox signaling, 2009Co-Authors: Hyun Ae Woo, Soo Han Bae, Sunjoo Park, Sue Goo RheeAbstract:Abstract The active-site Cysteine of 2-Cys peroxiredoxins (Prxs), a subgroup of the Prx family, is reversibly hyperoxidized to Cysteine Sulfinic Acid during catalysis with concomitant loss of peroxidase activity. The reduction of Sulfinic 2-Cys Prx enzymes, the first known biologic of such a reaction, has been reported to be catalyzed by either sulfiredoxin (Srx) or sestrin (Sesn) 2. The 13-kDa Srx and 60-kDa Sesn 2 show no sequence similarity, however. Whereas the reductase function of Srx has been confirmed by several studies, such is not the case for Sesn 2. We have now shown that (a) recombinant Sesn 2 did not catalyze the reduction of Sulfinic Prx I in vitro, whereas Srx did; (b) overexpression of Sesn 2 in HeLa or A549 cells did not affect the reduction of 2-Cys Prxs, whereas overexpression of Srx markedly increased the reduction rate; and (c) the rate of Sulfinic 2-Cys Prx reduction in embryonic fibroblasts derived from Sesn 2–knockout mice did not differ from that in those derived from wild-type m...
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sulfiredoxin the Cysteine Sulfinic Acid reductase specific to 2 cys peroxiredoxin its discovery mechanism of action and biological significance
Kidney International, 2007Co-Authors: Sue Goo Rhee, Woojin Jeong, Tong-shin ChangAbstract:Peroxiredoxin (Prx) is a family of bifunctional proteins that exhibit peroxidase and chaperone activities. Prx proteins contain a conserved Cys residue that undergoes a redox change between thiol and disulfide states. 2-Cys Prx enzymes, a subgroup of Prx family, are intrinsically susceptible to reversible hyperoxidation to Cysteine Sulfinic Acid during catalysis. Cysteine hyperoxidation of Prx was shown to result in loss of peroxidase activity and a concomitant gain of chaperone activity. Reduction of Sulfinic Prx enzymes, the first known biological example of such a reaction, is catalyzed by sulfiredoxin (Srx) in the presence of ATP. Srx appears to exist solely to support the reversible Sulfinic modification of 2-Cys Prx enzymes. Srx specifically binds to 2-Cys Prx enzymes by recognizing several critical surface-exposed residues of the Prxs, and transfer the γ -phosphate of ATP to their Sulfinic moiety, using its conserved Cysteine as the phosphate carrier. The resulting Sulfinic phosphoryl ester is reduced to Cysteine after oxidation of four thiol equivalents.
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Molecular Mechanism of the Reduction of Cysteine Sulfinic Acid of Peroxiredoxin to Cysteine by Mammalian Sulfiredoxin
The Journal of biological chemistry, 2006Co-Authors: Woojin Jeong, Sung Jun Park, Tong-shin Chang, Duck-yeon Lee, Sue Goo RheeAbstract:Abstract Among many proteins with Cysteine Sulfinic Acid (Cys-SO2H) residues, the Sulfinic forms of certain peroxiredoxins (Prxs) are selectively reduced by sulfiredoxin (Srx) in the presence of ATP. All Srx enzymes contain a conserved Cysteine residue. To elucidate the mechanism of the Srx-catalyzed reaction, we generated various mutants of Srx and examined their interaction with PrxI, their ATPase activity, and their ability to reduce Sulfinic PrxI. Our results suggest that three surface-exposed amino Acid residues, corresponding to Arg50, Asp57, and Asp79 of rat Srx, are critical for substrate recognition. The presence of the Sulfinic form (but not the reduced form) of PrxI induces the conserved Cysteine of Srx to take the γ-phosphate of ATP and then immediately transfers the phosphate to the Sulfinic moiety of PrxI to generate a Sulfinic Acid phosphoryl ester (Prx-Cys-S(=O)). This ester is reductively cleaved by a thiol molecule (RSH) such as GSH, thioredoxin, and dithiothreitol to produce a disulfide-S-monoxide (Prx-Cys-S(=O)-S-R). The disulfide-S-monoxide is further reduced through the oxidation of three thiol equivalents to complete the catalytic cycle and regenerate Prx-Cys-SH.
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reversing the inactivation of peroxiredoxins caused by Cysteine Sulfinic Acid formation
Science, 2003Co-Authors: Hyun Ae Woo, Kap Seok Yang, Sang-won Kang, Ho Zoon Chae, Sung Chul Hwang, Kanghwa Kim, Sue Goo RheeAbstract:The active-site Cysteine of peroxiredoxins is selectively oxidized to Cysteine Sulfinic Acid during catalysis, which leads to inactivation of peroxidase activity. This oxidation was thought to be irreversible. However, by metabolic labeling of mammalian cells with 35S, we show that the Sulfinic form of peroxiredoxin I, produced during the exposure of cells to H2O2, is rapidly reduced to the catalytically active thiol form. The mammalian cells9 ability to reduce protein Sulfinic Acid might serve as a mechanism to repair oxidatively damaged proteins or represent a new type of cyclic modification by which the function of various proteins is regulated.
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Reversible oxidation of the active site Cysteine of peroxiredoxins to Cysteine Sulfinic Acid. Immunoblot detection with antibodies specific for the hyperoxidized Cysteine-containing sequence
Journal of Biological Chemistry, 2003Co-Authors: Hyun Ae Woo, Kap Seok Yang, Hyung Ki Kim, Sang-won Kang, Ho Zoon Chae, Sue Goo RheeAbstract:We previously suggested that oxidation of the active site Cysteine of peroxiredoxin (Prx) I or Prx II to Cysteine Sulfinic Acid in H2O2-treated cells is reversible (Woo, H. A., Chae, H. Z., Hwang, S. C., Yang, K.-S., Kang, S. W., Kim, K., and Rhee, S. G. (2003) Science 300, 653-656). In contrast, it was recently proposed that sulfinylation of Prx II, but not that of Prx I or Prx III, is reversible (Chevallet, M., Wagner, E., Luche, S., van Dorssealaer, A., Leize-Wagner, E., and Rabilloud, T. (2003) J. Biol. Chem. 278, 37146-37153). The detection of sulfinylated proteins in both of these previous studies relied on complex proteomics analysis. We now describe a simple immunoblot assay for the detection of sulfinylated Prx enzymes that is based on antibodies produced in response to a sulfonylated peptide modeled on the conserved active site sequence. These antibodies recognized both Sulfinic and sulfonic forms of Prx equally well and allowed the detection of sulfinylated Prx enzymes in H2O2-treated cells with high sensitivity and specificity. With the use of these antibodies, we demonstrated that not only the cytosolic enzymes Prx I and Prx II but also the mitochondrial enzyme Prx III undergo reversible sulfinylation. The generation of antibodies specific for sulfonylated peptides should provide insight into protein function similar to that achieved with antibodies to peptides containing phosphoserine or phosphothreonine.
