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Ganglioside GM3 modulates conformation of reconstituted ca2+-ATPase

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Abstract

Using steady-state fluorescence and nanosecond time-resolved fluorescence techniques, the ca2+ ATPase conformational changes induced by ganglioside GM3 were studied with different quenchers. The results showed that GM3 could significantly increase the lifetime of intrinsic fluorescence of Ca2+-ATPase reconstituted into proteoliposomes, and could also weaken the intrinsic fluorescence quenching by KI or hypocrellin B, HB. Furthermore, by using quenching kinetic analysis of the time-resolved fluorescence, in the presence of GM3, the quenching constant (K3V) and quenching efficiency were significantly lowered. The obtained results suggest that the oligosaccharide chain and the ceramide moieties of the GM3 molecule could interact with its counterparts of the ca2+-ATPase respectively, thus change the conformation of the hydrophobic domain of the enzyme, making the tryptophan residues in different regions shift towards the hydrophilic-hydrophobic interface, and hence shorten the distance between the hydrophilic and the hydrophobic domains, making the enzyme with a more compact form exhibit higher enzyme activity.

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References

  1. Higashi, H., Yamagata, T., Mechanism for ganglioside-mediated modulation of a calmodulin-dependent enzyme,J. Biol. Chem., 1992, 267: 9839.

    PubMed  CAS  Google Scholar 

  2. Hakomori, S., Bifunctional role of glycosphingolipids,J. Biol. Chem., 1990, 265:18713.

    PubMed  CAS  Google Scholar 

  3. Wang, L. H., Tu, Y. P., Yang, X. Y.et al., Effect of ganglioside GM3 on the activity and conformation of reconstituted Ca2+-ATPase,FEBS Lett., 1996, 388:128.

    Article  PubMed  CAS  Google Scholar 

  4. Tu, Y. P., Yang, F. Y., Transmembrane ca2+ gradientmediated modulation of sarcoplasmic reticulum Ca2+-ATPase,Biochem. Biophys. Res. Commun., 1993, 196:561.

    Article  PubMed  CAS  Google Scholar 

  5. Tu, Y. P., Yang, F. Y., Fluorescence study on transmembrane ca2+ gradient-mediated conformation changes of sarcoplasmic reticulum Ca2+-ATPase,Bioscience Reports, 1994, 14:309.

    Article  PubMed  CAS  Google Scholar 

  6. Tsui, Z. C., Zhang, X. B., Tang, N. M.et al., Effect of a cell differentiation inducer ganglioside GM3 on the phospholipid metabolism of a human monocytoid leukemic J6-2 cells,Chinese Biochem. J., 1992, 8: 724.

    Google Scholar 

  7. Coll, R. J., Murphy, A. J., Purification of the ca2+-ATPase of sarcoplasmic reticulum by affinity chromatography,J. Biol. Chem., 1984, 259:14249.

    PubMed  CAS  Google Scholar 

  8. Gould, G. W., McWhirter, J. M., East, J. M.et al., Uptake of Ca2+ mediated by the Ca2+-Mg2+ ATPase in reconstituted vesicles,Biochim. Biophys. Acta, 1987, 904:36.

    Article  PubMed  CAS  Google Scholar 

  9. Yue, J. C., Tu, Y. P., Pang, S. Z., Studies of the tryptophan fluorescence quenching of sarcoplasmic reticulum ca2+-ATPase by hypocrellin B,Chinese Science Bulletin (in Chinese), 1995, 40:76.

    Google Scholar 

  10. Ferreira, S. T., Verjovski-Almeida, S., Fluorescence decay of sarcoplasmic reticulum ATPase: ligand binding and hydration effects,J. Biol. Chem., 1989, 264:15392.

    PubMed  CAS  Google Scholar 

  11. Ferreira, S. T., Fluorescence lifetime and quenching studies of sarcoplasmic reticulum Ca2+-adenosine-5-triphosphatase,Photochem. Photobiol., 1993, 58:195.

    Article  PubMed  CAS  Google Scholar 

  12. Inesi, G., Sumbilla, C., Kirtley, M. E., Relationships of molecular structure and function in ca2+-transport ATPase,Physiol. Rev., 1990, 70:749.

    PubMed  CAS  Google Scholar 

  13. Vilsen, B., Andersen, J. P., MacLennan, D. H., Functional consequences of alterations of hydrophobic amino acids located at the M4S4 boundary of the ca2+-ATPase of sarcoplasmic reticulum,J. Biol. Chem., 1991, 266:18839.

    PubMed  CAS  Google Scholar 

  14. de Meis, L., Martins, O. B., Alves, E. W., Role of water, hydrogen ion, and temperature on the synthesis of adenosine triphosphate by the sarcoplasmic reticulum adenosinetriphosphatase in the absence of a calcium gradient,Biochemistry, 1980, 19:4252.

    Article  PubMed  Google Scholar 

  15. de Meis, L., Role of water in the energy of hydrolysis of phosphate compounds—energy transduction in biological mern-branes,Biochim. Biophys. Acta, 1989, 973:333.

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Dalian Medical University, 116027, Dalian, China

    Lihua Wang & Zhaochun Cui

  2. State Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 100101, Beijing, China

    Xiaoyi Yang, Yaping Tu & Fuyu Yang

Authors
  1. Lihua Wang
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  2. Xiaoyi Yang
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  3. Yaping Tu
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  4. Zhaochun Cui
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  5. Fuyu Yang
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Project supported by the State Key Laboratory of Biomacromolecules.

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Wang, L., Yang, X., Tu, Y. et al. Ganglioside GM3 modulates conformation of reconstituted ca2+-ATPase. Sci. China Ser. C.-Life Sci. 40, 422–429 (1997). https://doi.org/10.1007/BF02881737

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  • DOI: https://doi.org/10.1007/BF02881737

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