The gain of rod phototransduction: reconciliation of biochemical and electrophysiological measurements |
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Authors: | Leskov I B Klenchin V A Handy J W Whitlock G G Govardovskii V I Bownds M D Lamb T D Pugh E N Arshavsky V Y |
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Institution: | Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA. |
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Abstract: | We have resolved a central and long-standing paradox in understanding the amplification of rod phototransduction by making direct measurements of the gains of the underlying enzymatic amplifiers. We find that under optimized conditions a single photoisomerized rhodopsin activates transducin molecules and phosphodiesterase (PDE) catalytic subunits at rates of 120-150/s, much lower than indirect estimates from light-scattering experiments. Further, we measure the Michaelis constant, Km, of the rod PDE activated by transducin to be 10 microM, at least 10-fold lower than published estimates. Thus, the gain of cGMP hydrolysis (determined by kcat/Km) is at least 10-fold higher than reported in the literature. Accordingly, our results now provide a quantitative account of the overall gain of the rod cascade in terms of directly measured factors. |
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