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Publication : Signaling states of rhodopsin in rod disk membranes lacking transducin βγ-complex.

First Author  Lomonosova E Year  2012
Journal  Invest Ophthalmol Vis Sci Volume  53
Issue  3 Pages  1225-33
PubMed ID  22266510 Mgi Jnum  J:196765
Mgi Id  MGI:5489861 Doi  10.1167/iovs.11-9350
Citation  Lomonosova E, et al. (2012) Signaling states of rhodopsin in rod disk membranes lacking transducin betagamma-complex. Invest Ophthalmol Vis Sci 53(3):1225-33
abstractText  PURPOSE: To characterize the possible role of transducin Gtbetagamma-complex in modulating the signaling properties of photoactivated rhodopsin and its lifetime in rod disc membranes and intact rods. METHODS: Rhodopsin photolysis was studied using UV-visible spectroscopy and rapid scanning spectroscopy in the presence of hydroxylamine in highly purified wild-type and Gtgamma-deficient mouse rod disc membranes. Complex formation between photoactivated rhodopsin and transducin was measured by extra-metarhodopsin (meta) II assay. Recovery of dark current and flash sensitivity in individual intact wild-type and Gtgamma-deficient mouse rods was measured by single-cell suction recordings. RESULTS: Photoconversion of rhodopsin to meta I/meta II equilibrium proceeds normally after elimination of the Gtbetagamma-complex. The meta I/meta II ratio, the rate of meta II decay, the reactivity of meta II toward hydroxylamine, and the rate of meta III formation in Gtgamma-deficient rod disc membranes were identical with those observed in wild-type samples. Under low-intensity illumination, the amount of extra-meta II in Gtgamma-deficient discs was significantly reduced. The initial rate of dark current recovery after 12% rhodopsin bleach was three times faster in Gtgamma-deficient rods, whereas the rate of the late current recovery was largely unchanged. Mutant rods also exhibited faster postbleach recovery of flash sensitivity. CONCLUSIONS: Photoactivation and thermal decay of rhodopsin proceed similarly in wild-type and Gtgamma-deficient mouse rods, but the complex formation between photoactivated rhodopsin and transducin is severely compromised in the absence of Gtbetagamma. The resultant lower transduction activation contributes to faster photoresponse recovery after a moderate pigment bleach in Gtgamma-deficient rods.
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