Light-induced activation of the rod phosphodiesterase leads to a rapid transient increase of near-infrared light scattering.
The so-called AT-signal described here is a transient light-induced increase of the near-infrared scattering from isolated bovine rod outer segments (ROS). Freshly prepared ROS are permeabilized with 0.01% Triton X-100 immediately before measurement in the presence of 1 mM GTP. The signal amplitude is saturated when approximately 2 rhodopsin molecules out of 30 000 are photo-excited. The signal recovers rapidly (approximately 90 s) and can be repeated in a succession of flashes. The AT-signal can be prevented by pre-activation of the phosphodiesterase (PDE) enzyme cascade at various levels: either at the level of G-protein, using ALF4- in darkness or GTP gamma S plus light; or at the level of the PDE catalytic unit, using protamine as an activator. The light sensitivity and kinetics of the AT-signal are similar to published parameters of PDE activation. These data suggest that light-induced activation of the PDE is the key reaction for the generation of the signal. On the other hand, blocking of the catalytic cGMP binding site by isobutylmethylxanthine only slightly affects the signal. We propose that the AT-signal reflects a structural change linked to the transient removal of the PDE inhibitory subunit from the catalytic unit.
K M
KM
Kamps
J
J
Reichert
K P
KP
Hofmann
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D006152:Cyclic GMP / Q000378:metabolism; D004789:Enzyme Activation / Q000528:radiation effects; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D007259:Infrared Rays; D008027:Light; D008565:Membrane Proteins / Q000378:metabolism; D010786:Photoreceptor Cells / Q000201:enzymology*; D011479:Protamines / Q000494:pharmacology; D012243:Rhodopsin / Q000378:metabolism / Q000528:radiation effects; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D012542:Scattering, Radiation; D013873:Thionucleotides / Q000494:pharmacology; D015293:Transducin
D008565:Membrane Proteins; D011479:Protamines; D013873:Thionucleotides; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D012243:Rhodopsin; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D015293:Transducin; D006152:Cyclic GMP; D015056:1-Methyl-3-isobutylxanthine