Deactivation Kinetics of the Transduction Cascade of Vision

The response of the retinal rod cell to a dim flash lasts less than a second. This phototransduction is mediated by a guanine nucleotide-binding (G) protein cascade in which rhodopsin is the receptor, transducin is the G-protein, and the cGMP-specific phosphodiesterase (PDE) is the effector. Photoex...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 1991-11, Vol.88 (21), p.9813-9817
Hauptverfasser: Vuong, T. Minh, Chabre, Marc
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Chabre, Marc
description The response of the retinal rod cell to a dim flash lasts less than a second. This phototransduction is mediated by a guanine nucleotide-binding (G) protein cascade in which rhodopsin is the receptor, transducin is the G-protein, and the cGMP-specific phosphodiesterase (PDE) is the effector. Photoexcited rhodopsin activates transducin which in turn activates PDE. For this underlying biochemistry to be kinetically compatible with the photoresponse, both transducin and PDE must be deactivated in subsecond times. We report here direct measurements of their deactivation kinetics. The rate of heat release when transducin and PDE hydrolyze, respectively, GTP and cGMP was measured using time-resolved microcalorimetry. With only GTP present, the heat pulse comes from the activation of transducin and its subsequent deactivation by endogenous GTP hydrolysis. The nonhydrolyzable analog guanine 5'-[γ-thio]triphosphate was used to distinguish between these two processes: about 40% of the total heat is due to activation. From the time course of the deactivation heat, the active lifetime of transducin is
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The nonhydrolyzable analog guanine 5'-[γ-thio]triphosphate was used to distinguish between these two processes: about 40% of the total heat is due to activation. From the time course of the deactivation heat, the active lifetime of transducin is &lt;1 s at 22⚬C. With both GTP and cGMP present, the highly amplified hydrolytic activity of the PDE is responsible for most of the heat produced; its rate of release is directly proportional to the amount of activated PDE. Measurements of this rate at low photoexcitation levels (e.g., 30 molecules of photoexcited rhodopsin per rod) provide much kinetic information about the cascade. Notably, deactivation of the PDE takes 0.6 s (at 23⚬C) and absolutely requires GTP hydrolysis. 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Minh</creatorcontrib><creatorcontrib>Chabre, Marc</creatorcontrib><title>Deactivation Kinetics of the Transduction Cascade of Vision</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>The response of the retinal rod cell to a dim flash lasts less than a second. This phototransduction is mediated by a guanine nucleotide-binding (G) protein cascade in which rhodopsin is the receptor, transducin is the G-protein, and the cGMP-specific phosphodiesterase (PDE) is the effector. Photoexcited rhodopsin activates transducin which in turn activates PDE. For this underlying biochemistry to be kinetically compatible with the photoresponse, both transducin and PDE must be deactivated in subsecond times. We report here direct measurements of their deactivation kinetics. The rate of heat release when transducin and PDE hydrolyze, respectively, GTP and cGMP was measured using time-resolved microcalorimetry. With only GTP present, the heat pulse comes from the activation of transducin and its subsequent deactivation by endogenous GTP hydrolysis. The nonhydrolyzable analog guanine 5'-[γ-thio]triphosphate was used to distinguish between these two processes: about 40% of the total heat is due to activation. From the time course of the deactivation heat, the active lifetime of transducin is &lt;1 s at 22⚬C. With both GTP and cGMP present, the highly amplified hydrolytic activity of the PDE is responsible for most of the heat produced; its rate of release is directly proportional to the amount of activated PDE. Measurements of this rate at low photoexcitation levels (e.g., 30 molecules of photoexcited rhodopsin per rod) provide much kinetic information about the cascade. Notably, deactivation of the PDE takes 0.6 s (at 23⚬C) and absolutely requires GTP hydrolysis. 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Psychology</topic><topic>Guanosine Triphosphate - metabolism</topic><topic>Heat</topic><topic>Hydrolysis</topic><topic>In Vitro Techniques</topic><topic>Kinetics</topic><topic>Molecular and cellular biology</topic><topic>Nucleotides</topic><topic>Photochemistry</topic><topic>Photoexcitation</topic><topic>Receptors</topic><topic>Rod Cell Outer Segment - metabolism</topic><topic>Signal reflection</topic><topic>Thermodynamics</topic><topic>transducin</topic><topic>Transducin - metabolism</topic><topic>Vision, Ocular</topic><topic>Vision, photoreception</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Vuong, T. 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For this underlying biochemistry to be kinetically compatible with the photoresponse, both transducin and PDE must be deactivated in subsecond times. We report here direct measurements of their deactivation kinetics. The rate of heat release when transducin and PDE hydrolyze, respectively, GTP and cGMP was measured using time-resolved microcalorimetry. With only GTP present, the heat pulse comes from the activation of transducin and its subsequent deactivation by endogenous GTP hydrolysis. The nonhydrolyzable analog guanine 5'-[γ-thio]triphosphate was used to distinguish between these two processes: about 40% of the total heat is due to activation. From the time course of the deactivation heat, the active lifetime of transducin is &lt;1 s at 22⚬C. With both GTP and cGMP present, the highly amplified hydrolytic activity of the PDE is responsible for most of the heat produced; its rate of release is directly proportional to the amount of activated PDE. Measurements of this rate at low photoexcitation levels (e.g., 30 molecules of photoexcited rhodopsin per rod) provide much kinetic information about the cascade. Notably, deactivation of the PDE takes 0.6 s (at 23⚬C) and absolutely requires GTP hydrolysis. This concurs with the subsecond lifetime of active transducin and means that, once GTP hydrolysis has occurred, the hitherto active PDE is quickly inhibited.</abstract><cop>Washington, DC</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>1658789</pmid><doi>10.1073/pnas.88.21.9813</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
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subjects 3',5'-Cyclic-GMP Phosphodiesterases - metabolism
3',5'-cyclic-nucleotide phosphodiesterase
Animals
Biochemistry
Biological and medical sciences
Biophysics
Calorimetry
Cattle
Cell physiology
Equilibrium flow
Fundamental and applied biological sciences. Psychology
Guanosine Triphosphate - metabolism
Heat
Hydrolysis
In Vitro Techniques
Kinetics
Molecular and cellular biology
Nucleotides
Photochemistry
Photoexcitation
Receptors
Rod Cell Outer Segment - metabolism
Signal reflection
Thermodynamics
transducin
Transducin - metabolism
Vision, Ocular
Vision, photoreception
title Deactivation Kinetics of the Transduction Cascade of Vision
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