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Thermal Denaturation of Proteins Studied by UV Spectroscopy

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Natasa Poklar and Gorazd Vesnaver

Faculty of Chemistry and Chemical Technology, University of Ljubljana, Askerceva 5, 1000 Ljubljana, Slovenia

J. Chem. Educ., 2000, 77 (3), p 380

DOI: 10.1021/ed077p380

Publication Date (Web): March 1, 2000

Abstract

UV spectroscopy has been widely used for monitoring the unfolding of proteins. During temperature-induced denaturation the protein absorbance changes with temperature until the process of unfolding is completed. Analysis of the measured absorbance-vs-temperature curve (UV melting curve) based on the two-state approximation of the denaturation process leads to the van't Hoff enthalpy of denaturation at the temperature of the half-transition, DH°(T_d), and the corresponding entropy of denaturation, DS°(T_d). A determination of DH°(T_d) and DS°(T_d) for a-chymotrypsinogen A at pH 3.0 from the experimental UV melting curve is demonstrated.

Keywords (Audience):

Second-Year Undergraduate

Keywords (Domain):

Analytical Chemistry

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Biophysical Chemistry

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