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Transition State Characterization of the Low- to Physiological-Temperature Nondenaturational Conformational Change in Bovine Adenosine Deaminase by Slow Scan Rate Differential Scanning Calorimetry

  • Bodnar, Melissa A. (Department of Chemistry and Physics, Texas Woman's University) ;
  • Britt, B. Mark (Department of Chemistry and Physics, Texas Woman's University)
  • Received : 2005.11.08
  • Accepted : 2005.12.09
  • Published : 2006.03.31
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Abstract

Bovine adenosine deaminase undergoes a nondenaturational conformational change at $29^{\circ}C$ upon heating which is characterized by a large increase in heat capacity. We have determined the transition state thermodynamics of the conformational change using a novel application of differential scanning calorimetry (DSC) which employs very slow scan rates. DSC scans at the conventional, and arbitrary, scan rate of $1^{\circ}C/min$ show no evidence of the transition. Scan rates from 0.030 to $0.20^{\circ}C/min$ reveal the transition indicating it is under kinetic control. The transition temperature $T_t$ and the transition temperature interval ${\Delta}T$ increase with scan rate. A first order rate constant $k_1$ is calculated at each $T_t$ from $k_1\;=\;r_{scan}/{\Delta}T$, where $r_{scan}$ is the scan rate, and an Arrhenius plot is constructed. Standard transition state analysis reveals an activation free energy ${\Delta}G^{\neq}$ of 88.1 kJ/mole and suggests that the conformational change has an unfolding quality that appears to be on the direct path to the physiological-temperature conformer.

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References

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