Reaction rate and collisional efficiency of the rhodopsin-transducin system in intact retinal rods.
A model of transducin activation is constructed from its partial reactions (formation of metarhodopsin II, association, and dissociation of the rhodopsin-transducin complex). The kinetic equations of the model are solved both numerically and, for small photoactivation, analytically. From data on the partial reactions in vitro, rate and activation energy profile of amplified transducin turnover are modeled and compared with measured light-scattering signals of transducin activation in intact retinal rods. The data leave one free parameter, the rate of association between transducin and rhodopsin. Best fit is achieved for an activation energy of 35 kJ/mol, indicating lateral membrane diffusion of the proteins as its main determinant. The absolute value of the association rate is discussed in terms of the success of collisions to form the catalytic complex. It is greater than 30% for the intact retina and 10 times lower after permeabilization with staphylococcus aureus alpha-toxin. Dissociation rates for micromolar guanosinetriphosphale (GTP) (Kohl, B., and K. P. Hofmann, 1987. Biophys. J. 52:271-277) must be extrapolated linearly up to the millimolar range to explain the rapid transducin turnover in situ. This is interpreted by an unstable rhodopsin-transducin-GTP transient state. At the time of maximal turnover after a flash, the rate of activation is determined as 30, 120, 800, 2,500, and 4,000 activated transducins per photoactivated rhodopsin and second at 5, 10, 20, 30, 37 degrees C, respectively.
Kahlert
M
M
Institut für Biophysik und Strahlenbiologie, Albert-Ludwigs-Universität, Freiburg, Federal Republic of Germany.
Hofmann
K P
KP
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D019204:GTP-Binding Proteins / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D008433:Mathematics; D008954:Models, Biological; D010786:Photoreceptor Cells / Q000502:physiology*; D012243:Rhodopsin / Q000378:metabolism*; D013816:Thermodynamics; D015293:Transducin / Q000378:metabolism*
D000244:Adenosine Diphosphate; D006160:Guanosine Triphosphate; D012243:Rhodopsin; D019204:GTP-Binding Proteins; D015293:Transducin