The Journal of General Physiology
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Published online
doi:10.1085/jgp.200810034
The Journal of General Physiology, Vol. 132, No. 5, 587-604
The Rockefeller University Press, 0022-1295 $30.00
© Astakhova et al.
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ARTICLE

Kinetics of Turn-offs of Frog Rod Phototransduction Cascade



Luba A. Astakhova, Michael L. Firsov, and Victor I. Govardovskii

Sechenov Institute for Evolutionary Physiology & Biochemistry, Russian Academy of Sciences, 194223 St. Petersburg, Russia

Correspondence to Michael L. Firstov: firsov{at}iephb.nw.ru

The time course of the light-induced activity of phototrandsuction effector enzyme cGMP-phosphodiesterase (PDE) is shaped by kinetics of rhodopsin and transducin shut-offs. The two processes are among the key factors that set the speed and sensitivity of the photoresponse and whose regulation contributes to light adaptation. The aim of this study was to determine time courses of flash-induced PDE activity in frog rods that were dark adapted or subjected to nonsaturating steady background illumination. PDE activity was computed from the responses recorded from solitary rods with the suction pipette technique in Ca2+-clamping solution.

A flash applied in the dark-adapted state elicits a wave of PDE activity whose rising and decaying phases have characteristic times near 0.5 and 2 seconds, respectively. Nonsaturating steady background shortens both phases roughly to the same extent. The acceleration may exceed fivefold at the backgrounds that suppress {approx}70% of the dark current.

The time constant of the process that controls the recovery from super-saturating flashes (so-called dominant time constant) is adaptation independent and, hence, cannot be attributed to either of the processes that shape the main part of the PDE wave. We hypothesize that the dominant time constant in frog rods characterizes arrestin binding to rhodopsin partially inactivated by phosphorylation. A mathematical model of the cascade that considers two-stage rhodopsin quenching and transducin inactivation can mimic experimental PDE activity quite well. The effect of light adaptation on the PDE kinetics can be reproduced in the model by concomitant acceleration on both rhodopsin phosphorylation and transducin turn-off, but not by accelerated arrestin binding. This suggests that not only rhodopsin but also transducin shut-off is under adaptation control.


Abbreviations used in this paper: GAP, GTP-ase–activating protein; IBMX, 3-isobutyl-1-methylxanthine; PDE, cGMP phosphodiesterase; ROS, rod outer segment.

© 2008 Astakhova et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jgp.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).


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