Evidence for cooperative Na+ and Cl− binding by strongly hydrated l-proline

In nature the amino acid L-proline (Pro) is a ubiquitous and highly effective osmolyte protecting cells against osmotic stress. To understand this effect knowledge of the hydration of Pro and its interactions with dissolved salts is essential. We studied these properties by combining statistical mechanics and broadband dielectric spectroscopy and found that Pro remains strongly hydrated up to high amino-acid concentrations. This is also the case upon NaCl addition to a 0.6 M Pro solution. Here, additionally a Pro center dot NaCl aggregate is formed with a stability constant of K degrees approximate to 0.95...1.25 M-1, where Na+ and Cl- cooperatively bind to adjacent carboxylate-oxygen and ammonium-hydrogen atoms, respectively