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Amide temperature coefficients in the protein G B1 domain
Jennifer H Tomlinson1, Mike P Williamson
1Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, UK.
Journal of Biomolecular NMR
|November 15, 2011
Summary
Temperature coefficients of protein G
Area of Science:
- Protein dynamics
- Nuclear magnetic resonance (NMR) spectroscopy
- Biophysical chemistry
Background:
- Protein G's B1 domain (GB1) is a model system for studying protein structure and dynamics.
- Temperature coefficients provide insights into molecular motion and stability.
- pH variations can significantly impact protein structure and dynamics.
Purpose of the Study:
- To investigate the temperature coefficients of backbone amide (1)H and (15)N nuclei in GB1.
- To determine the influence of temperature and pH on these coefficients.
- To correlate temperature coefficients with structural features and hydrogen bonding.
Main Methods:
- Nuclear magnetic resonance (NMR) spectroscopy was used to measure temperature coefficients.
- Experiments were conducted over a temperature range of 283–313 K.
- A pH range of 2.0 to 9.0 was explored to assess pH-dependent effects.
Main Results:
- Many nuclei exhibited pH-dependent temperature coefficients, with some fitted to one or two pK(a) values.
- (1)H coefficients showed that hydrogen-bonded amides have less negative values.
- The largest pH-dependent changes were observed in loop regions, suggesting temperature-dependent loss of structure is a key factor, rather than direct hydrogen bonding.
Conclusions:
- Temperature-dependent loss of structure, particularly in loop regions, significantly influences amide temperature coefficients.
- While hydrogen bonding affects (1)H coefficients, it is not the primary driver of pH-dependent changes.
- Amide (15)N coefficients did not show a clear relationship with protein structure in this study.
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