Ultrasonic studies of alcohol-induced transconformation in beta-lactoglobulin: the intermediate state.
In mixed alcohol-water solvents, bovine beta-lactoglobulin undergoes a cooperative transition from beta-sheet to a high alpha-helix content conformer. We report here the characterization of beta-lactoglobulin by compressibility and spectroscopy measurements during this transconformation. Both the volume and compressibility increase as a function of alcohol concentration, up to maximal values which depend on the chemical nature of the three alcohols used: hexafluoroisopropanol, trifluoroethanol, and isopropanol. The order of effectiveness of alcohols in inducing the compressibility transition is identical to that previously reported for circular dichroism and thus independent of the observation technique. The highly cooperative sigmoidal curves found by compressibility determination match closely those obtained by circular dichroism at 222 nm, indicating a correlation between the two phenomena measured by the two different techniques. The presence of an equilibrium intermediate form was shown by the interaction of beta-lactoglobulin with 8-anilino-1-naphthalene sulfonic acid, a probe widely used to detect molten-globule states of proteins. It was correlated with the plateau region of the volume curves and with the inflexion points of the sigmoidal compressibility curves. Ultrasound characterization of proteins can be carried out in optically transparent or nontransparent media.
Kanjilal
S
S
Laboratoire d'Imagerie Paramétrique, Université Pierre et Marie Curie, Paris, France.
Taulier
N
N
Le Huérou
J-Y
JY
Gindre
M
M
Urbach
W
W
Waks
M
M
D019840:2-Propanol / Q000494:pharmacology; D000438:Alcohols / Q000494:pharmacology*; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D002942:Circular Dichroism; D004305:Dose-Response Relationship, Drug; D007782:Lactoglobulins / Q000737:chemistry*; D020005:Propanols / Q000494:pharmacology; D011487:Protein Conformation; D017433:Protein Structure, Secondary / Q000187:drug effects; D014270:Trifluoroethanol / Q000494:pharmacology; D014465:Ultrasonics; D014466:Ultraviolet Rays; D006845:Hydrocarbons, Fluorinated
D019840:2-Propanol; D000438:Alcohols; D007782:Lactoglobulins; D020005:Propanols; D014270:Trifluoroethanol; C001337:hexafluoroisopropanol; D006845:Hydrocarbons, Fluorinated