Transient response of rod outer segment cGMP phosphodiesterase to actinic light pulses. II. Detailed quantitative kinetic model.
In the accompanying article (Schmidt, J.A., and Yguerabide, J. (1989) J. Biol. Chem. 264, 19790-19803), we presented a minimal quantitative kinetic model with one rate-limiting step for the transient response of rod outer segment (ROS) phosphodiesterase (PDE) to stimulating light pulses of low fractional bleach (linear response range) and showed that the model was in excellent quantitative agreement with experimental results. The model characterizes the PDE response in terms of the specific rate constant of the rate-limiting step, kL, the lifetime of photoactivated rhodopsin, tau R, and the lifetime of activated PDE, tau P, but makes no predictions on how these kinetic parameters should depend on the concentrations of the various reactive species involved in the PDE response to light and does not reveal the nature of the rate-limiting step. However, we established by curve fitting experimental data to theoretical expressions from the model that kL increases hyperbolically with [GTP], tau R decreases with [GTP], and tau P is independent of GTP. In this report we present three detailed kinetic models which make specific quantitative predictions on how the kinetic parameters of the minimal model should depend on nucleotide and G protein concentrations and test the models against experimental data. Each model consists of one rate-limiting step. The first detailed model postulates that the rate-limiting step is the dissociation of R*GT into R* and GT (T stands for GTP). The second model postulates that the rate-limiting step is the binding of GTP to R*G, and the third model postulates that the rate-limiting step is the encounter rate of R* and G on the ROS disc membrane. We find that only the first detailed model is consistent with the experimental results as characterized by the minimal model. Using this detailed model we (a) define kL and tau R in terms of more fundamental equilibrium and rate parameters, (b) develop a theory for the systematic evaluation of amplification or gain of the PDE light response from light-stimulated GTP-binding data as well as v(t) versus t graphs, and (c) clarify methods which have been used in the past to evaluate gain experimentally.
Yguerabide
J
J
Department of Biology, University of California, San Diego, La Jolla 92093.
Schmidt
J A
JA
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism / Q000528:radiation effects*; D000818:Animals; D002417:Cattle; D007700:Kinetics; D008027:Light; D008433:Mathematics; D008962:Models, Theoretical; D010786:Photoreceptor Cells / Q000201:enzymology*; D012374:Rod Cell Outer Segment / Q000201:enzymology*
D015106:3',5'-Cyclic-GMP Phosphodiesterases