NMR investigations on residue level unfolding thermodynamics in DLC8 dimer by temperature dependent native state hydrogen exchange |
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Authors: | P M Krishna Mohan Swagata Chakraborty Ramakrishna V Hosur |
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Institution: | (1) Department of Chemical Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Mumbai, 400 005, India |
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Abstract: | Understanding protein stability at residue level detail in the native state ensemble of a protein is crucial to understanding
its biological function. At the same time, deriving thermodynamic parameters using conventional spectroscopic and calorimetric
techniques remains a major challenge for some proteins due to protein aggregation and irreversibility of denaturation at higher
temperature values. In this regard, we describe here the NMR investigations on the conformational stabilities and related
thermodynamic parameters such as local unfolding enthalpies, heat capacities and transition midpoints in DLC8 dimer, by using
temperature dependent native state hydrogen exchange; this protein aggregates at high (>65°C) temperatures. The stability
(free energy) of the native state was found to vary substantially with temperature at every residue. Significant differences
were found in the thermodynamic parameters at individual residue sites indicating that the local environments in the protein
structure would respond differently to external perturbations; this reflects on plasticity differences in different regions
of the protein. Further, comparison of this data with similar data obtained from GdnHCl dependent native state hydrogen exchange
indicated many similarities at residue level, suggesting that local unfolding transitions may be similar in both the cases.
This has implications for the folding/unfolding mechanisms of the protein.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Dynein light chain protein Nuclear magnetic resonance Native state hydrogen exchange Thermal stability Unfolding energetics |
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