Amino Acids

, Volume 46, Issue 5, pp 1353–1365 | Cite as

Primary hepatocytes from mice lacking cysteine dioxygenase show increased cysteine concentrations and higher rates of metabolism of cysteine to hydrogen sulfide and thiosulfate

  • Halina Jurkowska
  • Heather B. Roman
  • Lawrence L. Hirschberger
  • Kiyoshi Sasakura
  • Tetsuo Nagano
  • Kenjiro Hanaoka
  • Jakub Krijt
  • Martha H. Stipanuk
Original Article

Abstract

The oxidation of cysteine in mammalian cells occurs by two routes: a highly regulated direct oxidation pathway in which the first step is catalyzed by cysteine dioxygenase (CDO) and by desulfhydration-oxidation pathways in which the sulfur is released in a reduced oxidation state. To assess the effect of a lack of CDO on production of hydrogen sulfide (H2S) and thiosulfate (an intermediate in the oxidation of H2S to sulfate) and to explore the roles of both cystathionine γ-lyase (CTH) and cystathionine β-synthase (CBS) in cysteine desulfhydration by liver, we investigated the metabolism of cysteine in hepatocytes isolated from Cdo1-null and wild-type mice. Hepatocytes from Cdo1-null mice produced more H2S and thiosulfate than did hepatocytes from wild-type mice. The greater flux of cysteine through the cysteine desulfhydration reactions catalyzed by CTH and CBS in hepatocytes from Cdo1-null mice appeared to be the consequence of their higher cysteine levels, which were due to the lack of CDO and hence lack of catabolism of cysteine by the cysteinesulfinate-dependent pathways. Both CBS and CTH appeared to contribute substantially to cysteine desulfhydration, with estimates of 56 % by CBS and 44 % by CTH in hepatocytes from wild-type mice, and 63 % by CBS and 37 % by CTH in hepatocytes from Cdo1-null mice.

Keywords

Cysteine Cysteine dioxygenase Cystathionine γ-lyase Cystathionine β-synthase Hydrogen sulfide Thiosulfate Mice Hepatocytes 

Notes

Acknowledgments

The authors thank Dr. John E. Dominy (Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA) for training us in the technique for murine hepatocyte isolation and Dr. Viktor Kožich (Institute of Inherited Metabolic Disorders, Charles University in Prague, First Faculty of Medicine and General University Hospital, Praha, Czech Republic) for facilitating the by LC–MS/MS analyses and reviewing the manuscript. This project was supported by Grant DK-056649 from the National Institute of Diabetes and Digestive and Kidney Diseases. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This study was supported in part by the research program of the Charles University in Prague (PRVOUK-P24/LF1/3) with access to the LC–MS/MS made possible by project OPPK No. CZ.2.16/3.1.00/24012. H.J. was supported by a “Mobility Plus” fellowship from the Ministry of Science and Higher Education (MNISW), Republic of Poland.

Conflict of interest

No conflict of interests, financial or otherwise, are reported by the authors.

References

  1. Abeles RH, Walsh CT (1973) Acetylenic enzyme inactivators. Inactivation of γ-cystathionase, in vitro and in vivo, by propargylglycine. J Am Chem Soc 95:6124–61251PubMedCrossRefGoogle Scholar
  2. Ang A, Konigstorfer A, Giles G, Bhatia M (2012) Measuring free tissue sulfide. Adv Biol Chem 2:360–365CrossRefGoogle Scholar
  3. Bella DL, Stipanuk MH (1995) Effects of protein, methionine, or chloride on acid-base balance and on cysteine catabolism. Am J Physiol 269:E910–E917PubMedGoogle Scholar
  4. Bradley H, Gough A, Sokhi RS, Hassell A, Waring R, Emery P (1994) Sulfate metabolism is abnormal in patients with rheumatoid arthritis. Confirmation by in vivo biochemical findings. J Rheumatol 21:1192–1196PubMedGoogle Scholar
  5. Brait M, Ling S, Nagpal JK, Chang X, Park HL, Lee J, Okamura J, Yamashita K, Sidransky D, Kim MS (2012) Cysteine dioxygenase 1 is a tumor suppressor gene silenced by promoter methylation in multiple human cancers. PLoS One 7(9):e44951PubMedCentralPubMedCrossRefGoogle Scholar
  6. Cavallini D, Mondovi B, De Marco C, Sciosciasantoro A (1962) Inhibitory effect of mercaptoethanol and hypotaurine on the desulfhydration of cysteine by cystathionase. Arch Biochem Biophys 96:456–457PubMedCrossRefGoogle Scholar
  7. Chen X, Jhee KH, Kruger WD (2004) Production of the neuromodulator H2S by cystathionine β-synthase via the condensation of cysteine and homocysteine. J Biol Chem 279:52082–52086PubMedCrossRefGoogle Scholar
  8. Chiku T, Padovani D, Zhu W, Singh S, Vitvitsky V, Banerjee R (2009) H2S biogenesis by human cystathionine γ-lyase leads to the novel sulfur metabolites lanthionine and homolanthionine and is responsive to the grade of hyperhomocysteinemia. J Biol Chem 284:11601–11612PubMedCentralPubMedCrossRefGoogle Scholar
  9. Cuevasanta E, Denicola A, Alvarez B, Möller MN (2012) Solubility and permeation of hydrogen sulfide in lipid membranes. PLoS ONE 7:e34562PubMedCentralPubMedCrossRefGoogle Scholar
  10. Czubak J, Wróbel M, Jurkowska H (2002) Cystathionine γ-lyase. An enzymatic assay of α-ketobutyrate using lactate dehydrogenase. Acta Biol Crac Ser Zool 44:113–117Google Scholar
  11. Di Meo I, Fagiolari G, Prelle A, Viscomi C, Zeviani M, Tiranti V (2011) Chronic exposure to sulfide causes accelerated degradation of cytochrome c oxidase in ethylmalonic encephalopathy. Antioxid Redox Signal 15:353–362PubMedCrossRefGoogle Scholar
  12. Dominy JE Jr, Hirschberger LL, Coloso RM, Stipanuk MH (2006) Regulation of cysteine dioxygenase degradation is mediated by intracellular cysteine levels and the ubiquitin-26 S proteasome system in the living rat. Biochem J 394:267–273PubMedCentralPubMedCrossRefGoogle Scholar
  13. Dominy JE Jr, Hwang J, Guo S, Hirschberger LL, Zhang S, Stipanuk MH (2008) Synthesis of amino acid cofactor in cysteine dioxygenase is regulated by substrate and represents a novel post-translational regulation of activity. J Biol Chem 283:12188–12201PubMedCentralPubMedCrossRefGoogle Scholar
  14. Dominy JE Jr, Lee Y, Jedrychowski MP, Chim H, Jurczak MJ, Camporez JP, Ruan HB, Feldman J, Pierce K, Mostoslavsky R, Denu JM, Clish CB, Yang X, Shulman GI, Gygi SP, Puigserver P (2012) The deacetylase Sirt6 activates the acetyltransferase GCN5 and suppresses hepatic gluconeogenesis. Mol Cell 48:900–913PubMedCentralPubMedCrossRefGoogle Scholar
  15. Dorman DC, Moulin FJ, McManus BE, Mahle KC, James RA, Struve MF (2002) Cytochrome oxidase inhibition induced by acute hydrogen sulfide inhalation: correlation with tissue sulfide concentrations in the rat brain, liver, lung, and nasal epithelium. Toxicol Sci 65:18–25PubMedCrossRefGoogle Scholar
  16. Hildebrandt TM, Grieshaber MK (2008) Three enzymatic activities catalyze the oxidation of sulfide to thiosulfate in mammalian and invertebrate mitochondria. FEBS J 275:3352–3361PubMedCrossRefGoogle Scholar
  17. Huang J, Khan S, O’Brien PJ (1998) The glutathione dependence of inorganic sulfate formation from l- or d-cysteine in isolated rat hepatocytes. Chem Biol Interact 110:189–202PubMedCrossRefGoogle Scholar
  18. Jurkowska H, Uchacz T, Roberts J, Wróbel M (2011) Potential therapeutic advantage of ribose-cysteine in the inhibition of astrocytoma cell proliferation. Amino Acids 41:131–139PubMedCrossRefGoogle Scholar
  19. Kabil O, Vitvitsky V, Xie P, Banerjee R (2011) The quantitative significance of the transsulfuration enzymes for H2S production in murine tissues. Antioxid Redox Signal 15:363–372PubMedCentralPubMedCrossRefGoogle Scholar
  20. Koj A, Frendo J, Janik Z (1967) [35S]thiosulphate oxidation by rat liver mitochondria in the presence of glutathione. Biochem J 103:791–795PubMedCentralPubMedGoogle Scholar
  21. Kolluru GK, Shen X, Bir SC, Kevil CG (2013) Hydrogen sulfide chemical biology: pathophysiological roles and detection. Nitric Oxide 35C:5–20CrossRefGoogle Scholar
  22. Kraus J, Packman S, Fowler B, Rosenberg LE (1978) Purification and properties of cystathionine β-synthase from human liver: improved purification scheme and additional characterization of the enzyme in crude and pure form. Arch Biochem Biophys 222:44–52CrossRefGoogle Scholar
  23. Lagoutte E, Mimoun S, Andriamihaja M, Chaumontet C, Blachier F, Bouillaud F (2010) Oxidation of hydrogen sulfide remains a priority in mammalian cells and causes reverse electron transfer in colonocytes. Biochim Biophys Acta 1797:1500–1511PubMedCrossRefGoogle Scholar
  24. Levitt MD, Abdel-Rehim MS, Furne J (2011) Free and acid-labile hydrogen sulfide concentrations in mouse tissues: anomalously high free hydrogen sulfide in aortic tissue. Antioxid Redox Signal 15:373–378PubMedCrossRefGoogle Scholar
  25. Linden DR, Furne J, Stoltz GJ, Abdel-Rehim MS, Levitt MD, Szurszewski JH (2012) Sulphide quinone reductase contributes to hydrogen sulphide metabolism in murine peripheral tissues but not in the CNS. Br J Pharmacol 165:2178–2190PubMedCentralPubMedCrossRefGoogle Scholar
  26. Matsuo Y, Greenberg DM (1958) A crystalline enzyme that cleaves homoserine and cystathionine. I. Isolation procedure and some physicochemical properties. J Biol Chem 230:545–560PubMedGoogle Scholar
  27. Nagy P, Pálinkás Z, Nagy A, Budai B, Tóth I, Vasas A (2014) Chemical aspects of hydrogen sulfide measurements in physiological samples. Biochim Biophys Acta 1840:876–891Google Scholar
  28. Nicholson RA, Roth SH, Zhang A, Zheng J, Brookes J, Skrajny B, Bennington R (1998) Inhibition of respiratory and bioenergetic mechanisms by hydrogen sulfide in mammalian brain. J Toxicol Environ Health A 54:491–507 (677:350–357)PubMedCrossRefGoogle Scholar
  29. Olson KR (2012) A practical look at the chemistry and biology of hydrogen sulfide. Antioxid Redox Signal 17:32–44PubMedCentralPubMedCrossRefGoogle Scholar
  30. Predmore BL, Lefer DJ, Gojon G (2012) Hydrogen sulfide in biochemistry and medicine. Antioxid Redox Signal 17:119–140PubMedCentralPubMedCrossRefGoogle Scholar
  31. Rao AM, Drake MR, Stipanuk MH (1990) Role of the transsulfuration pathway and of γ-cystathionase activity in the formation of cysteine and sulfate from methionine in rat hepatocytes. J Nutr 120:837–845PubMedGoogle Scholar
  32. Roman HB, Hirschberger LL, Krijt J, Valli A, Kožich V, Stipanuk MH (2013) The cysteine dioxgenase knockout mouse: altered cysteine metabolism in nonhepatic tissues leads to excess H2S/HS production and evidence of pancreatic and lung toxicity. Antioxid Redox Signal. doi:10.1089/ars.2012.5010 PubMedGoogle Scholar
  33. Sasakura K, Hanaoka K, Shibuya N, Mikami Y, Kimura Y, Komatsu T, Ueno T, Terai T, Kimura H, Nagano T (2011) Development of a highly selective fluorescence probe for hydrogen sulfide. J Am Chem Soc 133:18003–18005PubMedCrossRefGoogle Scholar
  34. Shibuya N, Tanaka M, Yoshida M, Ogasawara Y, Togawa T, Ishii K, Kimura H (2009) 3-Mercaptopyruvate sulfurtransferase produces hydrogen sulfide and bound sulfane sulfur in the brain. Antioxid Redox Signal 11:703–714PubMedCrossRefGoogle Scholar
  35. Shih VE, Carney MM, Mandell R (1979) A simple screening test for sulfite oxidase deficiency: detection of urinary thiosulfate by a modification of Sörbo’s method. Clin Chim Acta 95:143–145PubMedCrossRefGoogle Scholar
  36. Singh S, Padovani D, Leslie RA, Chiku T, Banerjee R (2009) Relative contributions of cystathionine β-synthase and γ-cystathionase to H2S biogenesis via alternative transsulfuration reactions. J Biol Chem 284:22457–22466PubMedCentralPubMedCrossRefGoogle Scholar
  37. Sörbo B (1957) A colorimetric method for the determination of thiosulfate. Biochim Biophys Acta 23:412–416PubMedCrossRefGoogle Scholar
  38. Stipanuk MH (1986) Metabolism of sulfur-containing amino acids. Annu Rev Nutr 6:179–209PubMedCrossRefGoogle Scholar
  39. Stipanuk MH (2004) Sulfur amino acid metabolism: pathways for production and removal of homocysteine and cysteine. Annu Rev Nutr 24:539–577PubMedCrossRefGoogle Scholar
  40. Stipanuk MH, Beck PW (1982) Characterization of the enzymic capacity for cysteine desulphhydration in liver and kidney of the rat. Biochem J 206:267–277PubMedCentralPubMedGoogle Scholar
  41. Stipanuk MH, Ueki I (2011) Dealing with methionine/homocysteine sulfur: cysteine metabolism to taurine and inorganic sulfur. J Inherit Metab Dis 34:17–32PubMedCentralPubMedCrossRefGoogle Scholar
  42. Stipanuk MH, Hirschberger LL, Londono MP, Cresenzi CL, Yu AF (2004) The ubiquitin-proteasome system is responsible for cysteine-responsive regulation of cysteine dioxygenase concentration in liver. Am J Physiol Endocrinol Metab 286:E439–E448PubMedCrossRefGoogle Scholar
  43. Stipanuk MH, Ueki I, Dominy JE Jr, Simmons CR, Hirschberger LL (2009) Cysteine dioxygenase: a robust system for regulation of cellular cysteine levels. Amino Acids 37:55–63PubMedCentralPubMedCrossRefGoogle Scholar
  44. Szczepkowski TW, Skarzynski B, Weber M (1961) The metabolic state of thiosulphate. Nature 189:1007–1008PubMedCrossRefGoogle Scholar
  45. Tiranti V, Viscomi C, Hildebrandt T, Di Meo I, Mineri R, Tiveron C, Levitt MD, Prelle A, Fagiolari G, Rimoldi M, Zeviani M (2009) Loss of ETHE1, a mitochondrial dioxygenase, causes fatal sulfide toxicity in ethylmalonic encephalopathy. Nat Med 15:200–205PubMedCrossRefGoogle Scholar
  46. Ueki I, Roman HB, Valli A, Fieselmann K, Lam J, Peters R, Hirschberger LL, Stipanuk MH (2011) Knockout of the murine cysteine dioxygenase gene results in severe impairment in ability to synthesize taurine and an increased catabolism of cysteine to hydrogen sulfide. Am J Physiol Endocrinol Metab 301:E668–E684PubMedCentralPubMedCrossRefGoogle Scholar
  47. Ueki I, Roman HB, Hirschberger LL, Junior CC, Stipanuk MH (2012) Extrahepatic tissues compensate for loss of hepatic taurine synthesis in mice with liver-specific knockout of cysteine dioxygenase. Am J Physiol Endocrinol Metab 302:E1292–E1299PubMedCentralPubMedCrossRefGoogle Scholar
  48. Uhteg LC, Westley J (1979) Purification and steady-state kinetic analysis of yeast thiosulfate reductase. Arch Biochem Biophys 195:211–222PubMedCrossRefGoogle Scholar
  49. Uren JR, Ragin R, Chaykovsky M (1978) Modulation of cysteine metabolism in mice—effects of propargylglycine and L-cyst(e)ine-degrading enzymes. Biochem Pharmacol 27:2807–2814PubMedCrossRefGoogle Scholar
  50. Valentine WN, Frankenfeld JK (1974) 3-Mercaptopyruvate sulfurtransferase (EC 2.8.1.2): a simple assay adapted to human blood cells. Clin Chim Acta 51:205–210PubMedCrossRefGoogle Scholar
  51. Washtien W, Abeles RH (1977) Mechanism of inactivation of γ-cystathionase by the acetylenic substrate analogue propargylglycine. Biochemistry 16:2485–2491PubMedCrossRefGoogle Scholar
  52. Whiteman M, Winyard PG (2011) Hydrogen sulfide and inflammation: the good, the bad, the ugly and the promising. Expert Rev Clin Pharmacol 4:13–32PubMedCrossRefGoogle Scholar
  53. Whiteman M, Le Trionnaire S, Chopra M, Fox B, Whatmore J (2011) Emerging role of hydrogen sulfide in health and disease: critical appraisal of biomarkers and pharmacological tools. Clin Sci (Lond) 121:459–488CrossRefGoogle Scholar
  54. Wróbel M, Jurkowska H, Sliwa L, Srebro Z (2004) Sulfurtransferases and cyanide detoxification in mouse liver, kidney, and brain. Toxicol Mech Methods 14:331–337PubMedCrossRefGoogle Scholar
  55. Yamanishi T, Tuboi S (1981) The mechanism of the L-cystine cleavage reaction catalyzed by rat liver γ-cystathionase. J Biochem 89:1913–1921PubMedGoogle Scholar
  56. Yang G, Wu L, Jiang B, Yang W, Qi J, Cao K, Meng Q, Mustafa AK, Mu W, Zhang S, Snyder SH, Wang R (2008) H2S as a physiologic vasorelaxant: hypertension in mice with deletion of cystathionine γ-lyase. Science 322:587–590PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • Halina Jurkowska
    • 1
    • 2
  • Heather B. Roman
    • 1
  • Lawrence L. Hirschberger
    • 1
  • Kiyoshi Sasakura
    • 3
  • Tetsuo Nagano
    • 3
  • Kenjiro Hanaoka
    • 3
  • Jakub Krijt
    • 4
  • Martha H. Stipanuk
    • 1
  1. 1.Division of Nutritional SciencesCornell UniversityIthacaUSA
  2. 2.Chair of Medical BiochemistryJagiellonian University Medical CollegeKrakówPoland
  3. 3.Graduate School of Pharmaceutical SciencesThe University of TokyoTokyoJapan
  4. 4.First Faculty of Medicine and General University Hospital, Institute of Inherited Metabolic DisordersCharles University in PraguePragueCzech Republic

Personalised recommendations