Decoupling of the Onset of Anharmonicity between a Protein and Its Surface Water around 200 K

  1. Institute of Natural Sciences, Shanghai Jiao Tong University, Shanghai 200240, China
  2. Department of Cell and Developmental Biology & Michigan Neuroscience Institute, University of Michigan Medical School, MI 48104, USA
  3. School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
  4. NIST Center for Neutron Research, National Institute of Standards and Technology (NIST), Gaithersburg, Maryland 20899, USA & Department of Materials Science and Engineering, University of Maryland, College Park, Maryland 20742, USA
  5. ISIS Pulsed Neutron and Muon Source, Rutherford Appleton Laboratory, Science & Technology Facilities Council, Didcot OX11 0QX, UK
  6. Neutron Science and Technology Center, Comprehensive Research Organization for Science and Society, 162-1 Shirakata, Tokai, Naka, Ibaraki 319-1106, Japan
  7. Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37931, USA

Peer review process

Not revised: This Reviewed Preprint includes the authors’ original preprint (without revision), an eLife assessment, and public reviews.

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Editors

  • Reviewing Editor
    Hannes Neuweiler
    University of Würzburg, Würzburg, Germany
  • Senior Editor
    Merritt Maduke
    Stanford University, Stanford, United States of America

Reviewer #1 (Public Review):

Summary:

Zheng et al. study the 'glass' transitions that occur in proteins at ca. 200K using neutron diffraction and differential isotopic labeling (hydrogen/deuterium) of the protein and solvent. To overcome limitations in previous studies, this work is conducted in parallel with 4 proteins (myoglobin, cytochrome P450, lysozyme, and green fluorescent protein) and experiments were performed at a range of instrument time resolutions (1ns - 10ps). The author's data looks compelling, and suggests that transitions in the protein and solvent behavior are not coupled and contrary to some previous reports, the apparent water transition temperature is a 'resolution effect'; i.e. instrument response is limited. This is likely to be important in the field, as a reassessment of solvent 'slaving' and the role of the hydration shell on protein dynamics should be reassessed in light of these findings.

Strengths:

The use of multiple proteins and instruments with a rate of energy resolution/ timescales.

Weaknesses:

The paper could be organised to better allow the comparison of the complete dataset collected.
The extent of hydration clearly influences the protein transition temperature. The authors suggest that "water can be considered here as lubricant or plasticizer which facilitates the motion of the biomolecule." This may be the case, but the extent of hydration may also alter the protein structure.

Reviewer #2 (Public Review):

Summary:

The manuscript entitled "Decoupling of the Onset of Anharmonicity between a Protein and Its Surface Water around 200 K" by Zheng et al. presents a neutron scattering study trying to elucidate if at the dynamical transition temperature water and protein motions are coupled. The origin of the dynamical transition temperature has been highly debated for decades, specifically its relation to hydration.

Strengths:

The study is rather well conducted, with a lot of effort to acquire the perdeuterated proteins, and some results are interesting.

Weaknesses:

The present work could certainly contribute some arguments, but I have the feeling that not all known facts are properly discussed.

The points the authors should carefully discuss are the following:

(1) Daniel et al. (10.1016/S0006-3495(98)77694-5) have shown that enzymes can be functional below the dynamical transition temperature which is at odds with some of the claims of the authors.

(2) It is not as easy to say that protonated proteins in D2O reflect protein dynamics while perdeuterated proteins in H2O reflect water dynamics. A recent study by Nidriche et al. (PRX LIFE 2, 013005 (2024)) reveals that H <-> D exchange is much faster than usually assumed and has important consequences for such studies.

(3) A publication by Jasnin et al. (10.1039/b923878f) on heparin sulfate shows a resolution effect.

(4) The authors should discuss the impact of the chosen q-range on their findings (see Phys. Chem. Chem. Phys., 2012, 14, 4927-4934, where the authors see a huge effect !).

(5) The authors underline that the dynamical transition is intrinsic to the protein. However, Cupane et al. (ref 12) have shown that it can also be found in a mixture of amino acids without any protein backbone.

(6) The authors say that they find similar dependences from MSD. They should explain that the MSD is inversely proportional to the summed intensities squared.

(7) A decoupling between water dynamics and membrane dynamics has already been discussed by K. Wood, G. Zaccai et al.

(8) The fact that transition temperature in lipid membranes is higher when the membrane is dry is also well known (A.V. Popova, D.K. Hincha, BMC Biophys. 4, 11 (2011)).

(9) The authors should mention the slope (K/min) they used for DSC and discuss the impact of it on the results.

(10) In the introduction, the authors should present the different explanations forwarded for the dynamical transition.

  1. Howard Hughes Medical Institute
  2. Wellcome Trust
  3. Max-Planck-Gesellschaft
  4. Knut and Alice Wallenberg Foundation