title: Concurrent observation of bulk and protein hydration water by spin-label ESR under nanoconfinement.
abstract: Under nanoconfinement the formation of crystalline ice is suppressed, allowing the study of water dynamics at subfreezing temperatures. Here we report a temperature-dependent investigation (170-260 K) of the behavior of hydration water under nanoconfinement by ESR techniques. A 26-mer-long peptide and the Bax protein are studied. This study provides site-specific information about the different local hydrations concurrently present in the protein/peptide solution, enabling a decent comparison of the hydration molecules-those that are buried inside, in contact with, and detached from the protein surface. Such a comparison is not possible without employing ESR under nanoconfinement. Though the confined bulk and surface hydrations behave differently, they both possess a transition similar to the reported fragile-to-strong crossover transition around 220 K. On the contrary, this transition is absent for the hydration near the buried sites of the protein. The activation energy determined under nanoconfinement is found to be lower in surface hydration than in bulk hydration. The protein structural flexibility, derived from the interspin distance distributions P(r) at different temperatures, is obtained by dipolar ESR spectroscopy. The P(r) result demonstrates that the structural flexibility is strongly correlated with the transition in the surface water, corroborating the origin of the protein dynamical transition at subfreezing temperatures.
authors.affiliation: Department of Chemistry and Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University , Hsinchu 30013, Taiwan.
authors.forename: Yun-Hsuan
authors.initials: YH
authors.lastname: Kuo
authors.forename: Yu-Ru
authors.initials: YR
authors.lastname: Tseng
authors.forename: Yun-Wei
authors.initials: YW
authors.lastname: Chiang
mesh_terms: D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D051379:Mice; D008958:Models, Molecular; D036103:Nanotechnology / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D013499:Surface Properties; D013696:Temperature; D014867:Water / Q000737:chemistry*; D051028:bcl-2-Associated X Protein / Q000737:chemistry*
chemical_list: D010455:Peptides; D051028:bcl-2-Associated X Protein; D014867:Water