Solvent effects on horse apomyoglobin dynamics.
The effects of the solvent conditions (buffer pH 9, 8, or 7 or buffer pH 6.5 alone or mixed with 3.2% ethanol or 6.2% formamide) on the protein dynamics of horse apomyoglobin were investigated through tryptophan fluorescence quenching, spectra, and decay properties. Raising the pH (which induces discontinuous protein conformation changes) increases the structural fluctuations inside the hydrophobic A, G, and H helix core. Mixed solutions containing either 3.2% ethanol or 6.2% formamide (which redistribute water molecules on the protein surface) produce protein dynamics changes in the vicinity of the two Trp residues, without inducing particular constraints on these very residues. Formamide increases, in the same way, the polarity and the protein flexibility while ethanol reduces both. The present fluorescence work also shows that, whatever the outside solvent, the two Trp residues W7 and W14, embedded in the A, G, and H helix core, are equally and statistically reached by small molecules diffusing inside the protein matrix. Hydrogen-tritium exchange measurements on the protein in mixed solvents reveal that the dynamics of the A, G, and H helix cluster and of the B and E helixes are greatly influenced by the nature of the outside medium. A small amount of formamide in the buffer increases the protein fluctuations while an ethanol-water mixture reduces them. We suggest that the hydratation state of the protein surface could be the relevant parameter of the protein dynamics.
Haouz
A
A
Laboratoire de Biologie Physico-Chimique, Universite Denis Diderot, 2 place Jussieu, 75251 Paris Cedex 05, France.
Glandieres
J M
JM
Zentz
C
C
Pin
S
S
Ramstein
J
J
Tauc
P
P
Brochon
J C
JC
Alpert
B
B
D000818:Animals; D001059:Apoproteins / Q000737:chemistry*; D006736:Horses; D008786:Metmyoglobin / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry*; D011487:Protein Conformation; D012997:Solvents; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry
D001059:Apoproteins; D009211:Myoglobin; D012997:Solvents; C010857:apomyoglobin; D008786:Metmyoglobin; D014364:Tryptophan