title: Effect of heavy water on protein flexibility.
abstract: The effects of heavy water (D(2)O) on internal dynamics of proteins were assessed by both the intrinsic phosphorescence lifetime of deeply buried Trp residues, which reports on the local structure about the triplet probe, and the bimolecular acrylamide phosphorescence quenching rate constant that is a measure of the average acrylamide diffusion coefficient through the macromolecule. The results obtained with several protein systems (ribonuclease T1, superoxide dismutase, beta-lactoglobulin, liver alcohol dehydrogenase, alkaline phosphatase, and apo- and Cd-azurin) demonstrate that in most cases D(2)O does significantly increase the rigidity the native structure. With the exception of alkaline phosphatase, the kinetics of the structure tightening effect of deuteration are rapid compared with the rate of H/D exchange of internal protons, which would then assign the dampening of structural fluctuations in D(2)O to a solvent effect, rather than to stronger intramolecular D bonding. Structure tightening by heavy water is generally amplified at higher temperatures, supporting a mostly hydrophobic nature of the underlying interaction, and under conditions that destabilize the globular fold.
authors.affiliation: Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Area della Ricerca di Pisa, Pisa, Italy. patrizia.cioni@ib.pi.cnr.it
authors.forename: Patrizia
authors.initials: P
authors.lastname: Cioni
authors.forename: Giovanni B
authors.initials: GB
authors.lastname: Strambini
mesh_terms: D020106:Acrylamide; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D017666:Deuterium Oxide / Q000737:chemistry*; D066298:In Vitro Techniques; D007700:Kinetics; D049449:Luminescence; D011487:Protein Conformation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D013816:Thermodynamics; D014364:Tryptophan / Q000031:analogs & derivatives*
chemical_list: D011506:Proteins; D020106:Acrylamide; C016952:N-acetyltryptophanamide; D014364:Tryptophan; D017666:Deuterium Oxide