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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Deuterium oxide stabilizes conformation of tubulin: a biophysical and biochemical study

  • Das, Amlan (Dr. B.C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta) ;
  • Sinha, Sharmistha (Molecular Biophysics Unit, Indian Institute Science) ;
  • Acharya, Bipul R (Dr. B.C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta) ;
  • Paul, Pinaki (Dr. B.C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta) ;
  • Bhattacharyya, Bhabatarak (Department of Biochemistry, Bose institute) ;
  • Chakrabarti, Gopal (Dr. B.C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta)
  • Published : 2008.01.31
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Abstract

The present study was aimed to elucidate the mechanism of stabilization of tubulin by deuterium oxide ($D_2O$). Rate of decrease of tryptophan fluorescence during aging of tubulin at 4$^{\circ}C$ and 37$^{\circ}C$ was significantly lower in $D_2O$ than in $H_2O$. Circular dichroism spectra of tubulin after incubation at 4$^{\circ}C$, suggested that complete stabilization of the secondary structure in D2O during the first 24 hours of incubation. The number of available cysteine measured by DTNB reaction was decreased to a lesser extent in $D_2O$ than in $H_2O$. . During the increase in temperature of tubulin, the rate of decrease of fluorescence at 335 nm and change of CD value at 222 nm was lesser in $D_2O$. Differential Scanning calorimetric experiments showed that the $T_m$ values for tubulin unfolding in $D_2O$ were 58.6$^{\circ}C$ and 62.17$^{\circ}C$, and in $H_2O$. those values were 55.4$^{\circ}C$ and 59.35$^{\circ}C$.

Keywords

References

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