Heat capacity analysis of oxidized Escherichia coli thioredoxin fragments (1–73, 74–108) and their noncovalent complex

Evidence for the burial of apolar surface in protein unfolded states

Authors


J. M. Sanchez-Ruiz, Facultad de Ciencias, Departamento de Quimica Fisica, Campus Fuentenueva s/n, 18071-Granada, Spain. Fax: + 34 958272879; Tel.: + 34 958243189; E-mail: sanchezr@goliat.ugr.es; M. L. Tasayco, Department of Chemistry, The City College of New York, Convent Avenue and 138 Street, New York, NY 10031, USA. Fax: + 1 212 6506107, Tel.: + 1 212 6501869, E-mail: mltj@libertad.sci.ccny.cuny.edu) or M. L. Tasayco, Department of Chemistry, The City College of New York, Convent Avenue and 138 Street, New York, NY 10031, USA. Tel.: + 1 212 6501869, Fax: + 1 212 6506107, E-mail: mltj@libertad.sci.ccny.cuny.edu.

Abstract

We have calculated the absolute heat capacities of fragments 1–73 (N fragment) and 74–108 (C fragment) from thioredoxin, their complex and the uncleaved protein, from the concentration dependence of the apparent heat capacities of the solutions determined by differential scanning calorimetry. We find that, while the absolute heat capacities of uncleaved, unfolded thioredoxin and the C fragment are in good agreement with the theoretical values expected for fully solvated chains (calculated as the sum of the contributions of the constituent amino acids), the absolute heat capacities of the N fragment and the unfolded complex are about 2 kJ·K−1·mol−1 lower than the fully solvated-chain values. We attribute this discrepancy to burial of the apolar surface in the N fragment (as burial of the polar area is expected to lead to an increase in heat capacity). Illustrative calculations suggest that burial of about 1000–1600 Å2 of apolar surface takes place in the N fragment (probably accompanied by the burial of a smaller amount of polar surface). In general, this work is supportive of heat capacity measurements on protein fragments being useful as probes of surface burial in studies to characterize protein unfolded states and the high regions of protein folding landscapes.

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