High Pressure NMR Reveals that Apomyoglobin is an Equilibrium Mixture from the Native to the Unfolded
Abstract
Pressure-induced reversible conformational changes of sperm whale apomyoglobin have been studied between 30 bar and 3000 bar on individual residue basis by utilizing 1H/15N hetero nuclear single-quantum coherence two-dimensional NMR spectroscopy at pH 6.0 and 35 degrees C. Apomyoglobin showed a series of pressure-dependent NMR spectra as a function of pressure, assignable to the native (N), intermediates (I), molten globule (MG) and unfolded (U) conformers. At 30 bar, the native fold (N) shows disorder only in the F helix. Between 500 bar and 1200 bar, a series of locally disordered conformers I are produced, in which local disorder occurs in the C helix, the CD loop, the G helix and part of the H helix. At 2000 bar, most cross-peaks exhibit severe line-broadening, suggesting the formation of a molten globule, but at 3000 bar all the cross-peaks reappear, showing that the molten globule turns into a well-hydrated, mobile unfolded conformation U. Since all the spectral changes were reversible with pressure, apomyoglobin is considered to exist as an equilibrium mixture of the N, I, MG and U conformers at all pressures. MG is situated at 2.4+/-(0.1) kcal/mol above N at 1 bar and the unfolding transition from the combined N-I state to MG is accompanied by a loss of partial molar volume by 75+/-(3) ml/mol. On the basis of these observations, we postulate a theorem that the partial molar volume of a protein decreases in parallel with the loss of its conformational order.
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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 [120]
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Wei, G., Xi, W., Nussinov, R., & Ma, B. (2016) · Chemical Reviews · doi:10.1021/acs.chemrev.5b00562 · Cited by 436 · see citing works
- Authors
- Kitahara, R., Yamada, H., Akasaka, K., & Wright, P. E.
- Affiliations
- Kobe University (JP)
- RIKEN (JP)
- Scripps Research Institute (US)
- Corresponding author
- Akasaka, Kazuyuki
- Venue
- Journal of Molecular Biology
- Publisher
- Elsevier BV
- Published
- 2002-07
- Volume
- 320
- Issue
- 2
- Pages
- 311-319
- ISSN
- 0022-2836 (ISSN-L)
- OpenAlex
- W1964038158
- Cited by
- 85
- References
- 51
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 2 · 2023: 5 · 2022: 1 · 2019: 4 · 2018: 2 · 2017: 5 · 2016: 3 · 2015: 6 · 2014: 4 · 2013: 5 · 2012: 7
- FWCI
- 1.97
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Ministry of Education, Culture, Sports, Science and Technology
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
- Physical Sciences › Chemistry › Spectroscopy › Advanced NMR Techniques and Applications
- Keywords
- apomyoglobin, sperm whale, high pressure, pressure-induced unfolding, molten globule, folding intermediates, conformational equilibrium, partial molar volume, protein hydration, line broadening, reversible conformational change
- MeSH terms
- Apoproteins, Escherichia coli, Models, Molecular, Myoglobin, Magnetic Resonance Spectroscopy, Protein Conformation, Thermodynamics, Protein Folding
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