Invited Symposium: SERCA-Type of Calcium Pumps and Phospholamban



Materials & Methods


Discussion & Conclusion



INABIS '98 Home Page Your Session Symposia & Poster Sessions Plenary Sessions Exhibitors' Foyer Personal Itinerary New Search

Topology of Catalytic, Ca2+ Binding and Thapsigargin Sites, and Allosteric Character of the SERCA ATPase

Contact Person: Giuseppe Inesi (ginesi@umaryland.edu)


  1. Autry, J. M. and Jones, L. R. (1997) Functional co-expression of the canine cardiac Ca2+ pump and phospholamban in Spodoptera frugiperda (Sf21) cells reveals new insights on ATPase regulation. Journal of Biological Chemistry, 272: 15872-15880.
  2. Bastide, F., Meissner, G., Fleischer, S., and Post, R. L. (1973) Similarity of the active site of phosphorylation of the ATPase for transport of sodium and potassium ions in kidney to that for transport of calcium ion in sarcoplasmic reticulum of muscle. Journal of Biological Chemistry, 248: 8385-8391.
  3. Bigelow, D. J. and Inesi, G. (1992) Contributions of Chemical Derivatization and Spectroscopic Studies to the Characterization of the Ca2+ Transport ATPase of Sarcoplasmic Reticulum. Biochimica et Biophysica Acta: Bio-Membranes, 1113: 323-338.
  4. Chiesi, M. and Inesi, G. (1980) Adenosine 5'-triphosphate dependent fluxes of manganese and hydrogen ions in sarcoplasmic reticulum. Biochemistry, 19: 2912-2918.
  5. Christensen, S. B., Andersen, A, and Smitt, UW. (1997) Sesquiterpenoids from Thapsia species and medicinal chemistry of the thapsigargins. Fortschr Chem Org Naturst, 71: 129-167.
  6. Clarke, D. M., Loo, T. W., Inesi, G., and MacLennan, D. H. (1989) Location of high affinity Ca2+-binding sites within the predicted transmembrane domain of the sarcoplasmic reticulum Ca2+-ATPase. Nature, 339: 476-478.
  7. Degani, C. and Boyer, P. D. (1973) A borohydride reduction method for characterization of the acyl phosphate linkage in proteins and its application to sarcoplasmic reticulum adenosine triphosphatase. Journal of Biological Chemistry, 248: 8222-8226.
  8. deMeis, L. and Vianna, A. (1979) Energy interconversion by the Ca2+-dependent ATPase of the sarcoplasmic reticulum. Annu.Rev.Biochem., 48: 275-292.
  9. Ho, S. N., Hunt, H. D., Horton, R. M., Pullen, J. K., and Pease, L. R. (1989) Site directed mutagenesis by overlap extension using the polymerase chain reaction. Gene, 77: 51-59.
  10. Hua, S., Malak, H., Lakowicz, J. R., and Inesi, G. (1995) Synthesis and Interaction of Fluorescent Thapsigargin Derivatives with the Sarcoplasmic Reticulum ATPase Membrane- Bound Region. Biochemistry, 34: 5137-5142.
  11. Hua, S. and Inesi, G. (1997) Synthesis of a radioactive azido derivative of thapsigargin and photolabeling of the sarcoplasmic reticulum ATPase. Biochemistry, 36(39): 11865-11872.
  12. Inesi, G. and Asai, H. (1968) Trypsin digestion of fragmented sarcoplasmic reticulum. Arch.Biochem.Biophys., 126: 469-477.
  13. Inesi, G. and Scales, D. J. (1974) Tryptic cleavage of sacroplasmic reticulum protein. Biochemistry, 13: 3299-3306.
  14. Inesi, G., Kurzmack, M., and Lewis, D. (1988) Kinetic and equilibrium characterization of an energy- transducing enzyme and its partial reactions. Methods Enzymol., 157: 154-190.
  15. Inesi, G., Sumbilla, C., and Kirtley, M. E. (1990) Relationships of Molecular Structure and Function in the Ca2+ Transport ATPase. Physiol.Rev., 70: 749-760.
  16. Inesi, G., Lewis, D., Nikic, D., and Kirtley, M. E. (1992) Long Range Intramolecular Linked Functions in the Calcium Transport ATPase. 185-215.
  17. Inesi, G. and Sagara, Y. (1994) Specific Inhibitors of Intracellular Ca2+ Transport ATPases. Journal of Membrane Biology, 141: 1-6.
  18. Karin, N. J., Kaprielian, Z., and Fambrough, D. M. (5-1989) Expression of avian Ca2+-ATPase in cultured mouse myogenic cells. Mol.Cell.Biol., 9((5)): 1978-1986.
  19. Lytton, J., Westlin, M., and Hanley, M. R. (1991) Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca-ATPase family of calcium pumps. Journal of Biological Chemistry, 266: 17067-17071.
  20. MacLennan, D. H., Brandl, C. J., Korczak, B., and Green, N. M. (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.
  21. Norregaard, A., Vilsen, B., and Andersen, J. P. (1993) Chimeric Ca2+-ATPase/Na+,K2+-ATPase molecules: Their phosphoenzyme intermediates and sensitivity to Ca2+ and thapsigargin. FEBS Lett., 336: 248-254.
  22. Ogawa, H., Stokes, D. L., Sasabe, H., and Toyoshima, C. (7-1998) Structure of the Ca2+ pump of sarcoplasmic reticulum: A view along the lipid bilayer at 9- resolution. Biophysical Journal, 75(1): 41-52.
  23. Sagara, Y. and Inesi, G. (1991) Inhibition of the sarcoplasmic reticulum Ca2+ transport ATPase by thapsigargin at subnanomolar concentrations. Journal of Biological Chemistry, 266: 13503-13506.
  24. Sagara, Y., Wade, J. B., and Inesi, G. (1992) A conformational mechanism for formation of a dead-end complex by the sarcoplasmic reticulum ATPase with thapsigargin. Journal of Biological Chemistry, 267: 1286-1292.
  25. Sagara, Y., Fernandez-Belda, F., deMeis, L., and Inesi, G. (1992) Characterization of the inhibition of intracellular Ca2+ transport ATPases by thapsigargin. Journal of Biological Chemistry, 267: 12606-12613.
  26. Strock, C., Cavagna, M., Peiffer, WE, Sumbilla, C., Lewis, D., and Inesi, G. (6-12-1998) Direct demonstration of Ca2+ binding defects in sarco-endoplasmic reticulum Ca2+ ATPase mutants overexpressed in COS-1 cells transfected with adenovirus vectors. Journal of Biological Chemistry, 273(24): 15104-15109.
  27. Sumbilla, C., Lu, L., Inesi, G., Ishii, T., Takeyasu, K., Fang, Y., and Fambrough, D. M. (1993) Ca2+ dependent and thapsigargin-inhibited phosphorylation of Na+,K+-ATPase catalytic domain following chimeric recombination with the Ca+2-ATPase. Journal of Biological Chemistry, 268: 21185-21192.
  28. Takebe, Y., Seiki, M., Fujisawa, J. I., Hoy, P., Yokota, K., Arai, K. I., Yoshida, M., and Arai, N. (1988) SRalpha Promoter: an efficient and versatile mammalian cDNA expression system composed of the Simian Virus 40 early promoter and the R-U5 segment of human T-Cell leukemia virus Type 1 long terminal repeat. Mol.Cell.Biol., 8: 466-472.
  29. Toyoshima, C., Sasabe, H., and Stokes, D. L. (1993) Three-dimensional cryo-electron microscopy of the calcium ion pump in the sarcoplasmic reticulum membrane. Nature, 362: 469-471.
  30. Witcome, M., Henderson, I., Lee, A. G., and East, J. M. (1992) Mechanism of the inhibition of the calcium pump of sarcoplasmic reticulum by thapsigargin. Biochemical Journal, 283(Pt. 2): 525-529.
  31. Yonekura, K, Stokes, D. L., Sasabe, H., and Toyoshima, C. (1997) The ATP-binding site of Ca(2+)-ATPase revealed by electron image Analysis. Biophys.J., 72(3): 997-1005.Yu, M., Zhong, L., Rishi, A. K., Khadeer, M., Inesi, G., and Hussain, A. (1998) Specific Substitutions at Amino Acid 256 of the Sarcoplasmic/Endoplasmic Reticulum Ca2+ Transport ATPase Mediate Resistance to Thapsigargin in Thapsigargin-resistant Hamster Cells. Journal of Biological Chemistry, 273(6): 3542-3546.
  32. Zhang, P, Toyoshima, C., Yonekura, K, Green, N. M., and Stokes, D. L. (4-23-1998) Structure of the calcium pump from sarcoplasmic reticulum at 8-A resolution. Nature, 392((6678)): 835-839.
  33. Zhang, Z., Sumbilla, C., Lewis, D., Summers, S., Klein, M. G., and Inesi, G. (1995) Mutational analysis of the peptide segment linking phosphyorylation and Ca2+-binding domains in the sarcoplasmic reticulum Ca2+-ATPase. Journal of Biological Chemistry, 270: 1-8.
  34. Zhong, L. and Inesi, G. (1998) Role of the S3 stalk segment in the thapsigargin concentration dependence of sarco-endoplasmic reticulum Ca2+-ATPase inhibition. Journal of Biological Chemistry, 273: 12994-12998.
  35. Acknowledgments: This work was supported by NIH Grant 2 PO1HL-27867. Manuscript edited by Jerry Domanico.

Back to the top.

<= Discussion & Conclussions REFERENCES

| Discussion Board | Next Page | Your Symposium |
Inesi, G.; Strock, C.; Zhong, L.; Kirtley, M.E.; (1998). Topology of Catalytic, Ca2+ Binding and Thapsigargin Sites, and Allosteric Character of the SERCA ATPase. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Invited Symposium. Available at URL http://www.mcmaster.ca/inabis98/wuytack/inesi0738/index.html
© 1998 Author(s) Hold Copyright