Subsecond deactivation of transducin by endogenous GTP hydrolysis.
The response of a retinal rod cell to a weak flash of light is mediated by a receptor/GTP-binding protein (rhodopsin/transducin) signal transduction system and terminates within a second. The T alpha subunit of transducin (composed of subunits T alpha, T beta and T gamma) is triggered by photoexcited rhodopsin (R*) to release GDP and bind GTP. The binding of GTP causes release of the T alpha unit from T beta gamma and allows it to modulate the activity of an enzyme that generates a second messenger. Termination of the response requires the hydrolysis of the GTP by intrinsic GTPase. As with other G proteins, the GTPase activity of transducin seems to be slow. Reported in vitro turnover rates of a few molecules of GTP hydrolysed per molecule of transducin per minute imply a T alpha-GTP deactivation time of many seconds. But this time might be only a small fraction of that of the GTPase cycle. We have now used time-resolved microcalorimetry in bovine rod outer segments (ROS) to monitor the heat release due to the hydrolysis of GTP by a transducin population that had been quickly activated by flash illumination of rhodopsin. The enthalpy of GTP hydrolysis is released within 1 s at 23 degrees C. This deactivation time seems to be independent of any diffusible factor in the preparation and concurs with the termination kinetics of the rod's response. Thereafter, transducin seems unable to reload GTP for many seconds. This refractory 'resetting' time may account for the low steady-state GTPase rates in vitro.
Vuong
T M
TM
Laboratoire de Biophysique Moléculaire et Cellulaire, (Unité Associée 520 du CNRS) Centre d'études Nucléaires de Grenoble, France.
Chabre
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D000818:Animals; D002417:Cattle; D006152:Cyclic GMP / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008027:Light; D010786:Photoreceptor Cells / Q000201:enzymology / Q000502:physiology*; D015398:Signal Transduction; D015293:Transducin / Q000502:physiology*; D014785:Vision, Ocular / Q000502:physiology*
D006160:Guanosine Triphosphate; D015293:Transducin; D006152:Cyclic GMP