title: Volume and enthalpy changes after photoexcitation of bovine rhodopsin: laser-induced optoacoustic studies.
abstract: Laser-induced optoacoustic measurements were performed with bovine rhodopsin in the temperature range 5-32 degrees C in its natural environment (i.e., in washed membranes) as well as solubilized in dodecyl-beta-D-maltoside. A signal deconvolution procedure using a simple sequential kinetic scheme for the photobaric time evolution revealed, in the case of the washed membranes, the presence of an intermediate with a 14-ns lifetime at 25 degrees C, of the same order as that reported for the BSI intermediate in solubilized rhodopsin (Hug, S. J., W. J. Lewis, C. M. Einterz, T. E. Thorgeirsson, and D. S. Kliger. 1990. Nanosecond photolysis of rhodopsin: evidence for a new, blue-shifted intermediate. Biochemistry. 29:1475-1485), with an energy content of (85 +/- 20) kJ/mol, and accompanied by an expansion of 26 +/- 3 ml/mol. The difference in energy content between BSI and the next transient lumi was estimated in only -1 +/- 5 kJ/mol, concomitant with an expansion of 9 +/- 3 ml/mol. Thus, this transition, which according to literature involves an equilibrium, should be controlled by an entropic change, rather than by an enthalpic difference. This is supported by the fact that both activation parameters for the decay of batho and BSI decrease upon solubilization. For detergent-solubilized rhodopsin, two time constants were enough to fit the sample signal. A short lifetime ascribable to BSI was not detected in this case. For the first intermediate (probably batho in equilibrium with BSI), an energy content of 50 +/- 20 kJ/mol and an expansion of 20 +/- 1 ml/mol, and for lumi an energy content of 11 +/- 20 kJ/mol and a further expansion of 11 +/- 2 ml/mol were determined. Thus, the intermediates of the membrane-embedded form of rhodopsin (in contrast to solubilized samples) are kept in a higher energy level, although the total expansion from rhodopsin to lumi is similar for both conditions (35 +/- 6 and 31 +/- 3 ml/mol). The expansions are interpreted as protein reorganization processes as a consequence of the photoisomerization of the chromophore. As a result, weak interactions are probably perturbed and the protein gains conformational flexibility.
authors.affiliation: Max-Planck-Institut für Strahlenchernie, Mülheim an der Ruhr, Germany.
authors.forename: J M
authors.initials: JM
authors.lastname: Strassburger
authors.forename: W
authors.initials: W
authors.lastname: Gärtner
authors.forename: S E
authors.initials: SE
authors.lastname: Braslavsky
mesh_terms: D000042:Absorption; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D007700:Kinetics; D007834:Lasers; D008956:Models, Chemical; D011487:Protein Conformation; D012243:Rhodopsin / Q000737:chemistry*; D013057:Spectrum Analysis; D013696:Temperature
chemical_list: D012243:Rhodopsin