Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC

Xiaoming He; Willem F. Wolkers; John H. Crowe; David J. Swanlund; John C Bischof

doi:10.1115/IMECE2002-33668

Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC

Xiaoming He
, Willem F. Wolkers
, John H. Crowe
, David J. Swanlund
, John C Bischof

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

2 Scopus citations

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 1/s for the FTIR study and 120.9kJ/mole and 3.6.10¹⁶ 1/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 1/s.

Original language	English (US)
Title of host publication	Advances in Heat and Mass Transfer in Biotechnology
Publisher	American Society of Mechanical Engineers (ASME)
Pages	39-43
Number of pages	5
ISBN (Print)	0791836398, 9780791836392
DOIs	https://doi.org/10.1115/IMECE2002-33668
State	Published - Jan 1 2002

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Name	ASME International Mechanical Engineering Congress and Exposition, Proceedings

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This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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10.1115/IMECE2002-33668

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He, X., Wolkers, W. F., Crowe, J. H., Swanlund, D. J., & Bischof, J. C. (2002). Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC. In Advances in Heat and Mass Transfer in Biotechnology (pp. 39-43). (ASME International Mechanical Engineering Congress and Exposition, Proceedings). American Society of Mechanical Engineers (ASME). https://doi.org/10.1115/IMECE2002-33668

Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC. / He, Xiaoming; Wolkers, Willem F.; Crowe, John H. et al.
Advances in Heat and Mass Transfer in Biotechnology. American Society of Mechanical Engineers (ASME), 2002. p. 39-43 (ASME International Mechanical Engineering Congress and Exposition, Proceedings).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

He, X, Wolkers, WF, Crowe, JH, Swanlund, DJ & Bischof, JC 2002, Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC. in Advances in Heat and Mass Transfer in Biotechnology. ASME International Mechanical Engineering Congress and Exposition, Proceedings, American Society of Mechanical Engineers (ASME), pp. 39-43. https://doi.org/10.1115/IMECE2002-33668

He X, Wolkers WF, Crowe JH, Swanlund DJ, Bischof JC. Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC. In Advances in Heat and Mass Transfer in Biotechnology. American Society of Mechanical Engineers (ASME). 2002. p. 39-43. (ASME International Mechanical Engineering Congress and Exposition, Proceedings). doi: 10.1115/IMECE2002-33668

He, Xiaoming ; Wolkers, Willem F. ; Crowe, John H. et al. / Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC. Advances in Heat and Mass Transfer in Biotechnology. American Society of Mechanical Engineers (ASME), 2002. pp. 39-43 (ASME International Mechanical Engineering Congress and Exposition, Proceedings).

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title = "Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC",

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 1/s for the FTIR study and 120.9kJ/mole and 3.6.1016 1/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 1/s.",

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AU - Crowe, John H.

AU - Swanlund, David J.

AU - Bischof, John C

PY - 2002/1/1

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N2 - 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 1/s for the FTIR study and 120.9kJ/mole and 3.6.1016 1/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 1/s.

AB - 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 1/s for the FTIR study and 120.9kJ/mole and 3.6.1016 1/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 1/s.

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T3 - ASME International Mechanical Engineering Congress and Exposition, Proceedings

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BT - Advances in Heat and Mass Transfer in Biotechnology

PB - American Society of Mechanical Engineers (ASME)

ER -

Investigation of in situ thermal denaturation of proteins in dunning AT-1 rat prostate cancer cells using FTIR and DSC

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