Mapping the Hydration Dynamics of Ubiquitin

• Published in: Journal of the American Chemical Society 2011-08, Vol.133 (32), p.12326-12329
• Main Authors: Nucci, Nathaniel V, Pometun, Maxim S, Wand, A. Joshua
• Format: Article
• Language: English
• Subjects: Humans; Models, Molecular; Nuclear Magnetic Resonance, Biomolecular - methods; Ubiquitin - chemistry; Water - chemistry
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja202033k

The nature of water’s interaction with biomolecules such as proteins has been difficult to examine in detail at atomic resolution. Solution NMR spectroscopy is potentially a powerful method for characterizing both the structural and temporal aspects of protein hydration but has been plagued by artifacts. Encapsulation of the protein of interest within the aqueous core of a reverse micelle particle results in a general slowing of water dynamics, significant reduction in hydrogen exchange chemistry and elimination of contributions from bulk water thereby enabling the use of nuclear Overhauser effects to quantify interactions between the protein surface and hydration water. Here we extend this approach to allow use of dipolar interactions between hydration water and hydrogens bonded to protein carbon atoms. By manipulating the molecular reorientation time of the reverse micelle particle through use of low viscosity liquid propane, the T1ρ relaxation time constants of 1H bonded to 13C were sufficiently lengthened to allow high quality rotating frame nuclear Overhauser effects to be obtained. These data supplement previous results obtained from dipolar interactions between the protein and hydrogens bonded to nitrogen and in aggregate cover the majority of the molecular surface of the protein. A wide range of hydration dynamics is observed. Clustering of hydration dynamics on the molecular surface is also seen. Regions of long-lived hydration water correspond with regions of the protein that participate in molecular recognition of binding partners suggesting that the contribution of the solvent entropy to the entropy of binding has been maximized through evolution.