Native state hydrogen exchange of cold shock protein A (CspA) has been characterized as a function of the denaturant urea and of the stabilizing agent trimethylamine N-oxide (TMAO). The structure of CspA has five strands of β-sheet. Strands β1–β4 have strongly protected amide protons that, based on experiments as a function of urea, exchange through a simple all-or-none global unfolding mechanism. By contrast, the protection of amide protons from strand β5 is too weak to measure in water. Strand β5 is hydrogen bonded to strands β3 and β4, both of which afford strong protection from solvent exchange. Gaussian network model (GNM) simulations, which assume that the degree of protection depends on tertiary contact density in the native structure, accurately predict the strong protection observed in strands β1–β4 but fail to account for the weak protection in strand β5. The most conspicuous feature of strand β5 is its low sequence hydrophobicity. In the presence of TMAO, there is an increase in the protection of strands β1–β4, and protection extends to amide protons in more hydrophilic segments of the protein, including strand β5 and the loops connecting the β-strands. TMAO stabilizes proteins by raising the free energy of the denatured state, due to highly unfavorable interactions between TMAO and the exposed peptide backbone. As such, the stabilizing effects of TMAO are expected to be relatively independent of sequence hydrophobicity. The present results suggest that the magnitude of solvent exchange protection depends more on solvent accessibility in the ensemble of exchange susceptible conformations than on the strength of hydrogen-bonding interactions in the native structure.