Cavity hydration as a gateway to unfolding: An NMR study of hen lysozyme at high pressure and low temperature
Abstract
We have used low temperatures (down to -20°C) and high pressures (up to 2000 bar) to populate low-lying excited state conformers of hen lysozyme, and have analyzed their structures site-specifically using (15)N/(1)H two-dimensional HSQC NMR spectroscopy. The resonances of a number of residues were found to be selectively broadened, as the temperature was lowered at a pressure of 2000 bar. The resulting disappearance of cross-peaks includes those of residues in the β-domain of the protein and the cleft between the β- and α-domains, both located close to water-containing cavities. The results indicate that low-lying excited state conformers of hen lysozyme are characterized by slowly fluctuating local conformations around these cavities, attributed to the opportunities for water molecules to penetrate into the cavities. Furthermore, we have found that these water-containing cavities are conserved in similar positions in lysozymes from a range of different biological species, indicating that they are a common evolutionary feature of this family of enzymes.
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Lower-volume conformations have their cavities occupied by solvent.
“In conformations with lower volume the cavities are filled with solvent.120–121”
From Protein Ensembles: How Does Nature Harness Thermodynamic Fluctuations for Life? The Diverse Functional Roles of Conformational Ensembles in the Cell (2016) · cites this paper as [121]
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- Authors
- Kamatari, Y. O., Smith, L. J., Dobson, C. M., & Akasaka, K.
- Affiliations
- Gifu University (JP)
- University of Oxford (GB)
- University of Cambridge (GB)
- Kindai University (JP)
- Corresponding author
- Akasaka, Kazuyuki
- Venue
- Biophysical Chemistry
- Publisher
- Elsevier BV
- Published
- 2011-06
- Volume
- 156
- Issue
- 1
- Pages
- 24-30
- ISSN
- 0301-4622 (ISSN-L)
- OpenAlex
- W2073445834
- Cited by
- 46
- References
- 57
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 1 · 2020: 2 · 2019: 1 · 2018: 2 · 2017: 4 · 2016: 5 · 2015: 5 · 2014: 4 · 2013: 8 · 2012: 6
- FWCI
- 2.59
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biophysical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- RIKEN
- Ministry of Education, Culture, Sports, Science and Technology (07F010)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Physical Sciences › Chemistry › Spectroscopy › Advanced NMR Techniques and Applications
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- hen egg white lysozyme, protein hydration, protein unfolding, low-temperature NMR, high pressure, beta-domain, protein folding, evolutionary conservation
- MeSH terms
- Animals, Chickens, Cold Temperature, Models, Molecular, Muramidase, Pressure, Protein Conformation, Water, Protein Structure, Tertiary, Nuclear Magnetic Resonance, Biomolecular, Protein Unfolding
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