Structural flexibility in hydrated proteins

The flexibility of protein structures is important in allowing the variety of motions, covering a wide range of magnitudes and frequencies, essential to biological activity. Protein flexibility is also implicated in denaturation, allowing proteins to take up nonactive conformations that have free energies close to that of the native state. High-frequency dielectric measurement can be used to study the flexibility of proteins by probing the relaxation of dipolar constituents of their structures. In this work, 14 hydrated globular proteins are investigated using this method. Four relaxation processes are identified, one of which, with a relaxation time of 19 ns, can be correlated with the sum of the number densities of protein glycine and alanine residues. A second with a relaxation time of 2 ns shows a dependence on the number of threonine residues. It is concluded that the dipolar peptide groups of the protein backbone associated with these residues are responsible for these dielectric responses, with the lower frequency dispersion being due to backbone mobility in the hydrophobic environment of the protein core and the higher frequency response being associated with mobility on the more hydrophilic protein surface. The correlation of protein backbone flexibility with particular side chains indicates that these protein motions are under the direct control of the amino acid sequence of the protein.