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Journal of Biomolecular NMR

, Volume 52, Issue 1, pp 57–64 | Cite as

Amide temperature coefficients in the protein G B1 domain

  • Jennifer H. Tomlinson
  • Mike P. Williamson
Article

Abstract

Temperature coefficients have been measured for backbone amide 1H and 15N nuclei in the B1 domain of protein G (GB1), using temperatures in the range 283–313 K, and pH values from 2.0 to 9.0. Many nuclei display pH-dependent coefficients, which were fitted to one or two pKa values. 1H coefficients showed the expected behaviour, in that hydrogen-bonded amides have less negative values, but for those amides involved in strong hydrogen bonds in regular secondary structure there is a negative correlation between strength of hydrogen bond and size of temperature coefficient. The best correlation to temperature coefficient is with secondary shift, indicative of a very approximately uniform thermal expansion. The largest pH-dependent changes in coefficient are for amides in loops adjacent to sidechain hydrogen bonds rather than the amides involved directly in hydrogen bonds, indicating that the biggest determinant of the temperature coefficient is temperature-dependent loss of structure, not hydrogen bonding. Amide 15N coefficients have no clear relationship with structure.

Keywords

Temperature coefficient Chemical shift Hydrogen bond pH Protein G 

Notes

Acknowledgments

We thank the Biotechnology and Biological Science Research Council (UK) for funding a studentship to JHT.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Department of Molecular Biology and BiotechnologyUniversity of SheffieldSheffieldUK
  2. 2.Institute of Molecular and Cellular BiologyUniversity of LeedsLeedsUK

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