A calorimetric study of solute effects on the kinetic stability of alpha-amylase

Søren Nymand Olsen, Kim Bruno Andersen, Lars Holm Øgendal, Peter Westh

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

In this study we evaluated the applications of isothermal titration calorimetry (ITC) to Study solute effects on the kinetics of irreversible protein denaturation. More specifically, denaturation of Bacillus Halmapalus alpha-amylase (BHA) was initiated by addition of EDTA to the calorimetric cell, which reduces the thermostability of the protein from marginally stable to unstable at the experimental temperature, by removing bound calcium ions. The calorimetric signal was shown to be proportional to the unfolding rate of the protein, since, Delta H-agg was shown to be close to zero. Comparison of ITC with chromatographic size exclusion data (SEC) provided an avenue to study unfolding and aggregation separately, which proved to be useful in analysing the mechanism of solute effects on denaturation kinetics. Solute effects are discussed in line with preferential interactions and Wyman linkage function
OriginalsprogEngelsk
TidsskriftThermochimica Acta
Vol/bind484
Udgave nummer1
Sider (fra-til)32-37
ISSN0040-6031
DOI
StatusUdgivet - 2009

Citer dette

Olsen, Søren Nymand ; Andersen, Kim Bruno ; Øgendal, Lars Holm ; Westh, Peter. / A calorimetric study of solute effects on the kinetic stability of alpha-amylase. I: Thermochimica Acta. 2009 ; Bind 484, Nr. 1. s. 32-37.
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abstract = "In this study we evaluated the applications of isothermal titration calorimetry (ITC) to Study solute effects on the kinetics of irreversible protein denaturation. More specifically, denaturation of Bacillus Halmapalus alpha-amylase (BHA) was initiated by addition of EDTA to the calorimetric cell, which reduces the thermostability of the protein from marginally stable to unstable at the experimental temperature, by removing bound calcium ions. The calorimetric signal was shown to be proportional to the unfolding rate of the protein, since, Delta H-agg was shown to be close to zero. Comparison of ITC with chromatographic size exclusion data (SEC) provided an avenue to study unfolding and aggregation separately, which proved to be useful in analysing the mechanism of solute effects on denaturation kinetics. Solute effects are discussed in line with preferential interactions and Wyman linkage function",
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A calorimetric study of solute effects on the kinetic stability of alpha-amylase. / Olsen, Søren Nymand; Andersen, Kim Bruno; Øgendal, Lars Holm; Westh, Peter.

I: Thermochimica Acta, Bind 484, Nr. 1, 2009, s. 32-37.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - A calorimetric study of solute effects on the kinetic stability of alpha-amylase

AU - Olsen, Søren Nymand

AU - Andersen, Kim Bruno

AU - Øgendal, Lars Holm

AU - Westh, Peter

PY - 2009

Y1 - 2009

N2 - In this study we evaluated the applications of isothermal titration calorimetry (ITC) to Study solute effects on the kinetics of irreversible protein denaturation. More specifically, denaturation of Bacillus Halmapalus alpha-amylase (BHA) was initiated by addition of EDTA to the calorimetric cell, which reduces the thermostability of the protein from marginally stable to unstable at the experimental temperature, by removing bound calcium ions. The calorimetric signal was shown to be proportional to the unfolding rate of the protein, since, Delta H-agg was shown to be close to zero. Comparison of ITC with chromatographic size exclusion data (SEC) provided an avenue to study unfolding and aggregation separately, which proved to be useful in analysing the mechanism of solute effects on denaturation kinetics. Solute effects are discussed in line with preferential interactions and Wyman linkage function

AB - In this study we evaluated the applications of isothermal titration calorimetry (ITC) to Study solute effects on the kinetics of irreversible protein denaturation. More specifically, denaturation of Bacillus Halmapalus alpha-amylase (BHA) was initiated by addition of EDTA to the calorimetric cell, which reduces the thermostability of the protein from marginally stable to unstable at the experimental temperature, by removing bound calcium ions. The calorimetric signal was shown to be proportional to the unfolding rate of the protein, since, Delta H-agg was shown to be close to zero. Comparison of ITC with chromatographic size exclusion data (SEC) provided an avenue to study unfolding and aggregation separately, which proved to be useful in analysing the mechanism of solute effects on denaturation kinetics. Solute effects are discussed in line with preferential interactions and Wyman linkage function

KW - Isothermal titration calorimetry

KW - Protein denaturation

KW - Preferential interactions

U2 - 10.1016/j.tca.2008.11.016

DO - 10.1016/j.tca.2008.11.016

M3 - Journal article

VL - 484

SP - 32

EP - 37

JO - Thermochimica Acta

JF - Thermochimica Acta

SN - 0040-6031

IS - 1

ER -