Abstract
The primary aim of the study was to test the influencesof thapsigargicin in releasing Ca2+ from the intracellular Ca2+ stores and comparing it with the effects of thapsigargin. In this study, the effect of tumour promoter thapsigargicin on intracellular Ca2+ has been described. Therefore, rat liver endoplasmic reticulum subcellular fractions (microsomes) were employed and Ca2+ movements measured by spectrofluorimeter. Fluo-3 studies were used to follow Ca2+ release in experiments with microsomes. When results are evaluated, it has been concluded that like thapsigargin, thapsigargicin has the ability of discharging the intracellular Ca2+ stores, increasing the intracellular free Ca2+ concentration ([Ca2+]i) and being a potent and specific inhibitor of the ER Ca2+-ATPase
Keywords
Ca2+ release intracellular stores SERCA ATPases spectroscopy thapsigargin (TG) thapsigargicin (TGC)
References
- Benech JC, Wolosker H, de Meis L (1995): Reversal of the Ca2+ pump of blood platelets. Biochem J, 306, 35-38.
- Carafoli E (2002): Calcium signaling: A tale for all seasons. PNAS, 99, 1115-1122.
- Carafoli E, Santella L, Branca D, Brini M (2001): Generation, Control, and Processing of Cellular Calcium Signals. Crit Rev Biochem Mol Bio, 36, 107-260.
- Caspersen C, Treiman M (1995): Thapsigargin discriminates strongly between Ca2+-ATPase phosphorylated intermediates with different subcellular distributions in bovine adrenal chromaffin cells. FEBS Lett, 377, 31-36.
- Christensen SB (1988): Interpretation of the NMR and circular dichroic data of the sesquiterpene lactone thapsigargin. Acta Chem Scand, B42, 623-628.
- Christensen SB, Andersen A, Poulsen JC, Treiman M (1993): Derivatives of thapsigargin as probes of its binding site on endoplasmic reticulum Ca2+ ATPase: Stereoselectivity and important functional groups. FEBS Lett, 335, 345-348.
- Cullen PJ, Irvine RF, Drobak BK, Dawson AP (1989): Inositol 1,3,4,5-tetrakisphosphate causes release of Ca from permeabilized mouse lymphoma L1210 cells by its conversion into inositol 1,4,5-trisphosphate. Biochem J, 259, 931-933.
- Dawson AP (1982): Kinetic properties of the Ca2+- accumulation system of a rat liver microsomal fraction. Biochem. J, 206, 73-79.
- Reinhart PH, Bygrave FL (1981): Glucagon stimulation of ruthenium red-insensitive calcium ion transport in developing rat liver. Biochem J, 194, 541-549.
- Dawson AP, Fulton DV (1983): Some properties of the Ca2+-stimulated ATPase of a rat liver microsomal fraction. Biochem J, 210, 405-410.
- Fiske CH, Subbarow Y (1925): The colorimetric determination of phosphorus. J Biol Chem, 66, 375-400.
- Goeger DE, Riley RT, Dorner JW, Cole RJ (1988): Cyclopiazonic acid inhibition of the Ca2+-transport ATPase in rat skeletal muscle sarcoplasmic reticulum vesicles. Biochem Pharmacol, 37, 978-981.
- Holzapfel CW (1968): The isolation and structure of cyclopiazonic acid, a toxic metabolite of penicillium cyclopium westling. Tetrahedron, 24, 2101-2119.
- Jackson TR, Patterson SI, Thastrup O, Hanley MR (1988): A novel tumour promoter, thapsigargin, transiently increases cytoplasmic free Ca2+ without generation of inositol phosphates in NG115-401L neuronal cells. Biochem J, 253, 81-86.
- Ley SV, Antonello A, Balskus EP, Booth DT, Christensen SB, Cleator E et al. (2004): Synthesis of the thapsigargins. PNAS, 101, 12073-12078.
- Lowry OH, Rosebrough NJ, Farr AL, Randal RJ (1951): Protein measurement with the folin phenol reagent. J Biol Chem, 248, 265-275.
- Lytton J, Westlin, M, Hanley MR (1991): Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca- ATPase family of calcium pumps. J Biol Chem, 266, 17067-17071.
- MacLennan DH, Brandl CJ, Korczak B, Green NM (1985): Amino-acid sequence of a Ca2+ + Mg2+ -dependent ATPase from rabbit muscle sarcoplasmic reticulum, deduced from its complementary DNA sequence. Nature, 316, 696-700.
- Moore GA, McConkey DJ, Kass GE, O’Brien PJ, Orrenius S (1987): 2,5-Di(tert-butyl)-1,4-benzohydroquinone - a novel inhibitor of liver microsomal Ca2+ sequestration. FEBS Lett, 224, 331-336.
- Murphy SJ, Schroeder RE, Blacker AM, Krasavage WJ, English JC (1992): A Study of Developmental Toxicity of Hydroquinone in the Rabbit. Fundam Appl Toxicol, 19, 214-221.
- Niggli V, Adunyah ES, Carafoli E (1981): Acidic phospholipids, unsaturated fatty acids, and limited proteolysis mimic the effect of calmodulin on the purified erythrocyte Ca2+-ATPase. J Biol Chem, 256, 8588-8592.
- Oztetik, E. (2009): Effects of Thapsigargicin on Ca Movements in L1210 Cells Permeabilized with Digitonin. Biochemistry-Moscow Suppl Series B: Biomed Chem, 3, 156-163.
- Papp B, Enyedi A, Kovacs T, Sarkadi B, Wuytack F, Thastrup O et al. (1991): Demonstration of two forms of calcium pumps by thapsigargin inhibition and radioimmunoblotting in platelet membrane vesicles. J Biol Chem, 266, 14593-14596.
- Rasmussen U, Christensen SB, Sandberg F (1978): Thapsigargin and thapsigargicin, two new histamin liberators from Thapsia garganica L. Acta Pharm Suec, 15, 133-140.
- Sagara, Y., Fernandez-Belda, F., de Meis, L. and Inesi, G. (1992): Characterization of the inhibition of intracellular Ca2+ transport ATPases by thapsigargin. J Biol Chem, 267, 12606-12613.
- Takemura H, Hughes AR, Thastrup O, Putney JW (1989): Activation of calcium entry by the tumor promoter thapsigargin in parotid acinar cells. Evidence that an intracellular calcium pool and not an inositol phosphate regulates calcium fluxes at the plasma membrane. J Biol Chem, 264, 12266-12271.
- Thastrup O, Cullen PJ, Drobak BK, Hanley MR, Dawson AP (1990): Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2+-ATPase. PNAS 87, 2466-2470.
- Thastrup O, Dawson AP, Scharff O, Foder B, Cullen PJ, Drobak BK (1989): Thapsigargin, a novel molecular probe for studying intracellular calcium release and storage. Agents Actions, 27, 17-23.
- Toyoshima C, Nakasako M, Nomura H, Ogawa H. (2000): Crystal structure of the calcium pump of sarcoplasmic reticulum at 2.6 Å resolution. Nature, 405, 647-655.
- Wictome M, Michelangeli F, Lee AG, East JM (1992): The inhibitors thapsigargin and 2,5-di(tert-butyl)-1,4- benzohydroquinone favour the E2 form of the Ca2+, Mg- ATPase. FEBS Lett, 304, 109-113.
Abstract
Bu çalışmanın ana amacı hücre içindeki Ca2+ depolarından Ca2+ salınımına tapsigargisinin etkisinin test edilmesi ve bunun tapsigargin etkisi ile karşılaştırılmasıdır. Çalışmada tümor promotörü tapsigargisinin hücre içindeki Ca2+’a etkileri tanımlanmıştır. Bu nedenle, sıçan endoplazmik retikulum subsellüler (hücrealtı) fraksiyonları (mikrozomlar) kullanılmış ve Ca2+hareketleri spektrofluorimetre ile ölçülmüştür. Mikrozomlardaki Ca2+ salınımı Fluo-3 çalışmaları ile takip edilmiştir. Elde edilen bulgular değerlendirildiğinde, tapsigargisinin de tapsigargin gibi hücre içi Ca2+ depolarını boşalttığı, hücre içi serbest Ca2+konsantrasyonunu ([Ca2+]i) arttırdığı ve ER Ca2+-ATPaz için güçlü ve özgül bir inhibitör olduğu kanısına varılmıştır
Keywords
Ca2+ salınımı hücreiçi depolar SERCA ATPaz spektroskopi tapsigargin (TG) tapsigargisin (TGC)
References
- Benech JC, Wolosker H, de Meis L (1995): Reversal of the Ca2+ pump of blood platelets. Biochem J, 306, 35-38.
- Carafoli E (2002): Calcium signaling: A tale for all seasons. PNAS, 99, 1115-1122.
- Carafoli E, Santella L, Branca D, Brini M (2001): Generation, Control, and Processing of Cellular Calcium Signals. Crit Rev Biochem Mol Bio, 36, 107-260.
- Caspersen C, Treiman M (1995): Thapsigargin discriminates strongly between Ca2+-ATPase phosphorylated intermediates with different subcellular distributions in bovine adrenal chromaffin cells. FEBS Lett, 377, 31-36.
- Christensen SB (1988): Interpretation of the NMR and circular dichroic data of the sesquiterpene lactone thapsigargin. Acta Chem Scand, B42, 623-628.
- Christensen SB, Andersen A, Poulsen JC, Treiman M (1993): Derivatives of thapsigargin as probes of its binding site on endoplasmic reticulum Ca2+ ATPase: Stereoselectivity and important functional groups. FEBS Lett, 335, 345-348.
- Cullen PJ, Irvine RF, Drobak BK, Dawson AP (1989): Inositol 1,3,4,5-tetrakisphosphate causes release of Ca from permeabilized mouse lymphoma L1210 cells by its conversion into inositol 1,4,5-trisphosphate. Biochem J, 259, 931-933.
- Dawson AP (1982): Kinetic properties of the Ca2+- accumulation system of a rat liver microsomal fraction. Biochem. J, 206, 73-79.
- Reinhart PH, Bygrave FL (1981): Glucagon stimulation of ruthenium red-insensitive calcium ion transport in developing rat liver. Biochem J, 194, 541-549.
- Dawson AP, Fulton DV (1983): Some properties of the Ca2+-stimulated ATPase of a rat liver microsomal fraction. Biochem J, 210, 405-410.
- Fiske CH, Subbarow Y (1925): The colorimetric determination of phosphorus. J Biol Chem, 66, 375-400.
- Goeger DE, Riley RT, Dorner JW, Cole RJ (1988): Cyclopiazonic acid inhibition of the Ca2+-transport ATPase in rat skeletal muscle sarcoplasmic reticulum vesicles. Biochem Pharmacol, 37, 978-981.
- Holzapfel CW (1968): The isolation and structure of cyclopiazonic acid, a toxic metabolite of penicillium cyclopium westling. Tetrahedron, 24, 2101-2119.
- Jackson TR, Patterson SI, Thastrup O, Hanley MR (1988): A novel tumour promoter, thapsigargin, transiently increases cytoplasmic free Ca2+ without generation of inositol phosphates in NG115-401L neuronal cells. Biochem J, 253, 81-86.
- Ley SV, Antonello A, Balskus EP, Booth DT, Christensen SB, Cleator E et al. (2004): Synthesis of the thapsigargins. PNAS, 101, 12073-12078.
- Lowry OH, Rosebrough NJ, Farr AL, Randal RJ (1951): Protein measurement with the folin phenol reagent. J Biol Chem, 248, 265-275.
- Lytton J, Westlin, M, Hanley MR (1991): Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca- ATPase family of calcium pumps. J Biol Chem, 266, 17067-17071.
- MacLennan DH, Brandl CJ, Korczak B, Green NM (1985): Amino-acid sequence of a Ca2+ + Mg2+ -dependent ATPase from rabbit muscle sarcoplasmic reticulum, deduced from its complementary DNA sequence. Nature, 316, 696-700.
- Moore GA, McConkey DJ, Kass GE, O’Brien PJ, Orrenius S (1987): 2,5-Di(tert-butyl)-1,4-benzohydroquinone - a novel inhibitor of liver microsomal Ca2+ sequestration. FEBS Lett, 224, 331-336.
- Murphy SJ, Schroeder RE, Blacker AM, Krasavage WJ, English JC (1992): A Study of Developmental Toxicity of Hydroquinone in the Rabbit. Fundam Appl Toxicol, 19, 214-221.
- Niggli V, Adunyah ES, Carafoli E (1981): Acidic phospholipids, unsaturated fatty acids, and limited proteolysis mimic the effect of calmodulin on the purified erythrocyte Ca2+-ATPase. J Biol Chem, 256, 8588-8592.
- Oztetik, E. (2009): Effects of Thapsigargicin on Ca Movements in L1210 Cells Permeabilized with Digitonin. Biochemistry-Moscow Suppl Series B: Biomed Chem, 3, 156-163.
- Papp B, Enyedi A, Kovacs T, Sarkadi B, Wuytack F, Thastrup O et al. (1991): Demonstration of two forms of calcium pumps by thapsigargin inhibition and radioimmunoblotting in platelet membrane vesicles. J Biol Chem, 266, 14593-14596.
- Rasmussen U, Christensen SB, Sandberg F (1978): Thapsigargin and thapsigargicin, two new histamin liberators from Thapsia garganica L. Acta Pharm Suec, 15, 133-140.
- Sagara, Y., Fernandez-Belda, F., de Meis, L. and Inesi, G. (1992): Characterization of the inhibition of intracellular Ca2+ transport ATPases by thapsigargin. J Biol Chem, 267, 12606-12613.
- Takemura H, Hughes AR, Thastrup O, Putney JW (1989): Activation of calcium entry by the tumor promoter thapsigargin in parotid acinar cells. Evidence that an intracellular calcium pool and not an inositol phosphate regulates calcium fluxes at the plasma membrane. J Biol Chem, 264, 12266-12271.
- Thastrup O, Cullen PJ, Drobak BK, Hanley MR, Dawson AP (1990): Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2+-ATPase. PNAS 87, 2466-2470.
- Thastrup O, Dawson AP, Scharff O, Foder B, Cullen PJ, Drobak BK (1989): Thapsigargin, a novel molecular probe for studying intracellular calcium release and storage. Agents Actions, 27, 17-23.
- Toyoshima C, Nakasako M, Nomura H, Ogawa H. (2000): Crystal structure of the calcium pump of sarcoplasmic reticulum at 2.6 Å resolution. Nature, 405, 647-655.
- Wictome M, Michelangeli F, Lee AG, East JM (1992): The inhibitors thapsigargin and 2,5-di(tert-butyl)-1,4- benzohydroquinone favour the E2 form of the Ca2+, Mg- ATPase. FEBS Lett, 304, 109-113.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA85RV66KP |
| Journal Section | Research Article |
| Authors | |
| Publication Date | June 1, 2012 |
| Published in Issue | Year 2012Volume: 59 Issue: 2 |