Nutritional Regulation of Cystein Dioxygenase

Summary

Principal Investigator: MARTHA STIPANUK
Affiliation: Cornell University
Country: USA
Abstract: Maintenance of a low cellular cysteine level is essential for cellular integrity, but having a sufficiently high cellular cysteine level to ensure adequate rates of synthesis of glutathione, coenzyme A, and proteins is also critical. Hepatic cysteine dioxygenase (CDO) activity plays a central role in regulating the partitioning of cysteine to meet various metabolic demands while at the same time maintaining low cysteine levels in the body by disposing of excess cysteine. Hepatic CDO activity increases more than 30-fold within hours after rats are switched from a 10% protein diet to a 40% protein diet, while hepatic cysteine levels remain less than 0.1 mmol/g. This upregulation of CDO is largely due to decreased ubiquitination and degradation of CDO by the 26S proteasome. The inhibition of CDO polyubiquitination can be effected by cysteamine, as well as cysteine, in cultured hepatocyte systems, suggesting cysteine itself may be the regulatory molecule. Evidence of abnormal or deficient CDO activity, including elevated cysteine and low sulfate concentrations, has been reported in individuals with a variety of diseases, both non-neurological and neurological, suggesting heterogeneity in CDO expression in the human population and a role of CDO activity in the etiology of several chronic diseases associated with aging. The major goal of this project is to further elucidate the molecular mechanisms involved in the marked changes in CDO levels that occur in response to dietary protein or SAAs. The specific aims for the proposed work are: (a.) To further characterize the two isoforms of CDO, the processes involved in their formation, and their relative enzymatic activity. (b.) To determine the physical structure of CDO and to elucidate the catalytic mechanism and details of the active site structure as well as sites and conformations involved in the action of cysteine in protecting CDO from rapid degradation. (c.) To evaluate the role of protein degradation, in particular the ubiquitin-proteasome pathway, in the regulation of the level of expressed CDO and to elucidate the role of cysteine in the regulation of CDO degradation. (d.) To evaluate the physiological significance of CDO in the regulation of cellular cysteine (and gtutathione) level.
Funding Period: 2000-03-01 - 2009-03-31
more information: NIH RePORT

Top Publications

  1. pmc Upregulation of capacity for glutathione synthesis in response to amino acid deprivation: regulation of glutamate-cysteine ligase subunits
    Angelos K Sikalidis
    Division of Nutritional Sciences, Cornell University, Ithaca, NY, 14853, USA
    Amino Acids 46:1285-96. 2014
  2. pmc Cysteine dioxygenase structures from pH4 to 9: consistent cys-persulfenate formation at intermediate pH and a Cys-bound enzyme at higher pH
    Camden M Driggers
    Department of Biochemistry and Biophysics, 2011 Ag and Life Sciences Building, Oregon State University, Corvallis, OR 97331, USA
    J Mol Biol 425:3121-36. 2013
  3. pmc The cysteine dioxgenase knockout mouse: altered cysteine metabolism in nonhepatic tissues leads to excess H2S/HS(-) production and evidence of pancreatic and lung toxicity
    Heather B Roman
    1 Division of Nutritional Sciences, Cornell University, Ithaca, New York
    Antioxid Redox Signal 19:1321-36. 2013
  4. pmc Linking crystallographic model and data quality
    P Andrew Karplus
    Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA
    Science 336:1030-3. 2012
  5. pmc Dealing with methionine/homocysteine sulfur: cysteine metabolism to taurine and inorganic sulfur
    Martha H Stipanuk
    Division of Nutritional Sciences, Cornell University, Ithaca, NY, 14853, USA
    J Inherit Metab Dis 34:17-32. 2011
  6. pmc Thiol dioxygenases: unique families of cupin proteins
    Martha H Stipanuk
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Amino Acids 41:91-102. 2011
  7. pmc Growing rats respond to a sulfur amino acid-deficient diet by phosphorylation of the alpha subunit of eukaryotic initiation factor 2 heterotrimeric complex and induction of adaptive components of the integrated stress response
    Angelos K Sikalidis
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    J Nutr 140:1080-5. 2010
  8. pmc Gene expression and integrated stress response in HepG2/C3A cells cultured in amino acid deficient medium
    Angelos K Sikalidis
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Amino Acids 41:159-71. 2011
  9. pmc 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
    Iori Ueki
    Division of Nutritional Sciences, Cornell University, Ithaca, New York 14853, USA
    Am J Physiol Endocrinol Metab 301:E668-84. 2011
  10. pmc Extrahepatic tissues compensate for loss of hepatic taurine synthesis in mice with liver-specific knockout of cysteine dioxygenase
    Iori Ueki
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Am J Physiol Endocrinol Metab 302:E1292-9. 2012

Scientific Experts

  • MARTHA STIPANUK
  • John E Dominy
  • Iori Ueki
  • Lawrence L Hirschberger
  • Chad R Simmons
  • Angelos K Sikalidis
  • Heather B Roman
  • P Andrew Karplus
  • Jeong In Lee
  • Camden M Driggers
  • Alessandro Valli
  • Tadhg P Begley
  • Relicardo M Coloso
  • Quan Hao
  • Kevin M Mazor
  • Richard B Cooley
  • Jakub Krijt
  • Viktor Kozich
  • Banumathi Sankaran
  • Kay Diederichs
  • Carolyn Junior
  • Jimmy Lam
  • Rachel Peters
  • Krista Fieselmann
  • David J Schuller
  • Spencer L Granett
  • Kalyanaraman Krishnamoorthy
  • Qun Liu
  • Mari S Machi
  • Qingqiu Huang

Detail Information

Publications25

  1. pmc Upregulation of capacity for glutathione synthesis in response to amino acid deprivation: regulation of glutamate-cysteine ligase subunits
    Angelos K Sikalidis
    Division of Nutritional Sciences, Cornell University, Ithaca, NY, 14853, USA
    Amino Acids 46:1285-96. 2014
    ..Upregulation of GCLM levels in response to low cysteine levels may serve to protect the cell in the face of a future stress requiring GSH as an antioxidant or conjugating/detoxifying agent. ..
  2. pmc Cysteine dioxygenase structures from pH4 to 9: consistent cys-persulfenate formation at intermediate pH and a Cys-bound enzyme at higher pH
    Camden M Driggers
    Department of Biochemistry and Biophysics, 2011 Ag and Life Sciences Building, Oregon State University, Corvallis, OR 97331, USA
    J Mol Biol 425:3121-36. 2013
    ..5 and ~7.5 that influence Cys binding and oxygen binding/reactivity, respectively. Furthermore, this work provides reliable ligand-bound models for guiding future mechanistic considerations. ..
  3. pmc The cysteine dioxgenase knockout mouse: altered cysteine metabolism in nonhepatic tissues leads to excess H2S/HS(-) production and evidence of pancreatic and lung toxicity
    Heather B Roman
    1 Division of Nutritional Sciences, Cornell University, Ithaca, New York
    Antioxid Redox Signal 19:1321-36. 2013
    ....
  4. pmc Linking crystallographic model and data quality
    P Andrew Karplus
    Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA
    Science 336:1030-3. 2012
    ..CC* also can be used to assess model and data quality on the same scale, and this reveals when data quality is limiting model improvement...
  5. pmc Dealing with methionine/homocysteine sulfur: cysteine metabolism to taurine and inorganic sulfur
    Martha H Stipanuk
    Division of Nutritional Sciences, Cornell University, Ithaca, NY, 14853, USA
    J Inherit Metab Dis 34:17-32. 2011
    ..Cysteine dioxygenase is robustly regulated in response to cysteine availability and may function to provide a pathway to siphon cysteine to less toxic metabolites than those produced by cysteine desulfuration reactions...
  6. pmc Thiol dioxygenases: unique families of cupin proteins
    Martha H Stipanuk
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Amino Acids 41:91-102. 2011
    ..In ADOs, the conserved glutamate residue in cupin motif 1 is replaced by either glycine or valine. Both ADOs and CDOs appear to represent unique clades within the cupin superfamily...
  7. pmc Growing rats respond to a sulfur amino acid-deficient diet by phosphorylation of the alpha subunit of eukaryotic initiation factor 2 heterotrimeric complex and induction of adaptive components of the integrated stress response
    Angelos K Sikalidis
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    J Nutr 140:1080-5. 2010
    ..Taken together, our results indicate that rats fed the SAA-deficient diet had a prolonged activation of an eIF2alpha kinase that leads to upregulation of adaptive components of the ISR...
  8. pmc Gene expression and integrated stress response in HepG2/C3A cells cultured in amino acid deficient medium
    Angelos K Sikalidis
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Amino Acids 41:159-71. 2011
    ..Based on genes highly differentially expressed in both leucine- and cysteine-deficient cells, a list of 67 downregulated and 53 upregulated genes is suggested as likely targets of essential amino acid deprivation in mammalian cells...
  9. pmc 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
    Iori Ueki
    Division of Nutritional Sciences, Cornell University, Ithaca, New York 14853, USA
    Am J Physiol Endocrinol Metab 301:E668-84. 2011
    ..Control of cysteine levels by regulation of CDO may be necessary to maintain low H(2)S/sulfane sulfur levels and facilitate the use of H(2)S as a signaling molecule...
  10. pmc Extrahepatic tissues compensate for loss of hepatic taurine synthesis in mice with liver-specific knockout of cysteine dioxygenase
    Iori Ueki
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Am J Physiol Endocrinol Metab 302:E1292-9. 2012
    ....
  11. pmc 3T3-L1 adipocytes and rat adipose tissue have a high capacity for taurine synthesis by the cysteine dioxygenase/cysteinesulfinate decarboxylase and cysteamine dioxygenase pathways
    Iori Ueki
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    J Nutr 139:207-14. 2009
    ..Overall, our results demonstrate that CDO is regulated at both the level of enzyme abundance and the level of mRNA in mature adipocytes...
  12. pmc Cysteine dioxygenase: a robust system for regulation of cellular cysteine levels
    M H Stipanuk
    Division of Nutritional Sciences, Cornell University, 227 Savage Hall, Ithaca, NY 14853, USA
    Amino Acids 37:55-63. 2009
    ....
  13. pmc A putative Fe2+-bound persulfenate intermediate in cysteine dioxygenase
    Chad R Simmons
    Division of Nutritional Sciences, Cornell University, Ithaca, New York 14853, USA
    Biochemistry 47:11390-2. 2008
    ..A nonpolar environment of the distal oxygen would facilitate isomerization of the persulfenate to the sulfinate product...
  14. ncbi Expression, purification, and kinetic characterization of recombinant rat cysteine dioxygenase, a non-heme metalloenzyme necessary for regulation of cellular cysteine levels
    Chad R Simmons
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853 6301, USA
    Protein Expr Purif 47:74-81. 2006
    ..45 mM for L-cysteine. The expression and purification of CDO in a stable, highly active form has yielded significant insight into the kinetic properties of this unique thiol dioxygenase...
  15. pmc Preparation, crystallization and X-ray diffraction analysis to 1.5 A resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation
    Chad R Simmons
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853 8001, USA
    Acta Crystallogr Sect F Struct Biol Cryst Commun 61:1013-6. 2005
    ..55, c = 123.06 A, alpha = beta = gamma = 90 degrees. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initio phasing using a selenomethionyl derivative...
  16. ncbi Crystal structure of mammalian cysteine dioxygenase. A novel mononuclear iron center for cysteine thiol oxidation
    Chad R Simmons
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853 8001, USA
    J Biol Chem 281:18723-33. 2006
    ....
  17. ncbi Mammalian cysteine metabolism: new insights into regulation of cysteine metabolism
    Martha H Stipanuk
    Division of Nutritional Sciences, Cornell University, Ithaca, NY, USA
    J Nutr 136:1652S-1659S. 2006
    ....
  18. pmc Identification and characterization of bacterial cysteine dioxygenases: a new route of cysteine degradation for eubacteria
    John E Dominy
    Division of Nutritional Sciences, 227 Savage Hall, Cornell University, Ithaca, NY 14853, USA
    J Bacteriol 188:5561-9. 2006
    ..Suggestions are made for how this novel pathway of cysteine metabolism may play a role in the life cycle of the eubacteria that have it...
  19. ncbi Cysteamine dioxygenase: evidence for the physiological conversion of cysteamine to hypotaurine in rat and mouse tissues
    Relicardo M Coloso
    Cornell University, Division of Nutritional Sciences, Ithaca, New York 148453, USA
    Adv Exp Med Biol 583:25-36. 2006
  20. ncbi In vivo regulation of cysteine dioxygenase via the ubiquitin-26S proteasome system
    John E Dominy
    Cornell University, Division of Nutritional Sciences, Ithaca, 14853 New York, USA
    Adv Exp Med Biol 583:37-47. 2006
  21. ncbi Overexpression of cysteine dioxygenase reduces intracellular cysteine and glutathione pools in HepG2/C3A cells
    John E Dominy
    Department of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Am J Physiol Endocrinol Metab 293:E62-9. 2007
    ..WT CDO also decreased the glutathione pool and potentiated the toxicity of CdCl(2). These results demonstrate that CDO is capable of altering intracellular cysteine levels as well as glutathione levels...
  22. ncbi Discovery and characterization of a second mammalian thiol dioxygenase, cysteamine dioxygenase
    John E Dominy
    Department of Nutritional Sciences, Cornell University, Ithaca, New York 14853, USA
    J Biol Chem 282:25189-98. 2007
    ..Overall, these data suggest that ADO is responsible for endogenous cysteamine dioxygenase activity...
  23. pmc Enzymes of the taurine biosynthetic pathway are expressed in rat mammary gland
    Iori Ueki
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    J Nutr 137:1887-94. 2007
    ....
  24. pmc Regulation of cysteine dioxygenase degradation is mediated by intracellular cysteine levels and the ubiquitin-26 S proteasome system in the living rat
    John E Dominy
    Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA
    Biochem J 394:267-73. 2006
    ..The present study represents the first demonstration of regulated ubiquitination and degradation of a protein in a living mammal, inhibition of which had dramatic effects on cysteine catabolism...