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Investigation of In Situ Thermal Denaturation of Proteins in Dunning AT-1 Rat Prostate Cancer Cells Using FTIR and DSC

[+] Author Affiliations
Xiaoming He, David J. Swanlund, John C. Bischof

University of Minnesota, Minneapolis, MN

Willem F. Wolkers, John H. Crowe

University of California at Davis, Davis, CA

Paper No. IMECE2002-33668, pp. 39-43; 5 pages
doi:10.1115/IMECE2002-33668
From:
  • ASME 2002 International Mechanical Engineering Congress and Exposition
  • Advances in Heat and Mass Transfer in Biotechnology
  • New Orleans, Louisiana, USA, November 17–22, 2002
  • Conference Sponsors: Heat Transfer Division
  • ISBN: 0-7918-3639-8 | eISBN: 0-7918-1691-5, 0-7918-1692-3, 0-7918-1693-1
  • Copyright © 2002 by ASME

abstract

In this study, the in situ protein denaturation in Dunning AT-1 rat prostate cells was studied using FTIR and DSC. The denaturation information from FTIR showed a shift from α helix (amide-I band at 1655cm−1 ) to β sheet (amide I band at 1620 cm−1 ). The relative beta sheet area change between 20 °C and 70 °C was used to dynamically scale the thermal denaturation process during heating at 2 °C/min. DSC scans (at scanning rates of 2°C/min and 5°C/min) of the heat flow due to protein denaturation were recorded from 20 °C to 70 °C. The range over which protein denaturation occurs (from 45 °C to 70 °C) was consistent in both studies. By calorimetric experiments with DSC, the enthalpy change during protein denaturation was found to be 27.3±4.0J/g J/g and 25.9±0.5J/g, respectively. In addition, the activation energy and frequency factor were measured by kinetic experiments with both DSC and FTIR. A first order irreversible Arrhenius model was fit to the kinetic data using a flexible tolerance method. The activation energy (E) and frequency factor (A) between 20–70 °C was found to be 107.8kJ/mole and 3.4·1014 l/s for the FTIR study and 120.9kJ/mole and 3.6·1016 l/s for DSC analysis at a scanning rate of 2°C/min, respectively. At a scanning rate of 5°C/min, the DSC results gave an activation energy of 144.5kJ/mole and a frequency factor of 4.1·1020 l/s.

Copyright © 2002 by ASME

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