Light-induced interaction between rhodopsin and the GTP-binding protein. Metarhodopsin II is the major photoproduct involved.
We have previously described [H, Kühn et al. (1981) Proc. Natl Acad. Sci. USA, 78, 6873-6877] a light-induced scattering change ('binding signal') associated with a stoichiometric binding between photoexcited rhodopsin and a peripheral membrane protein, the GTP-binding protein, in bovine rod outer segment suspensions. We have attempted here to identify the rhodopsin intermediate R* which is responsible for this interaction, by studying its dependence on pH, temperature and ionic strength. The results strongly suggest that the active state is metarhodopsin II (M II). 1. The initial phase of the binding signal is slightly slower than the formation of metarhodopsin II (2-37 degrees C, pH 5.5-9). 2. The kinetics of the decay of the active rhodopsin state are similar to those of the metarhodopsin II leads to metarhodopsin III transition (37 degrees C, pH 7.3). 3. All conditions which lead to light-induced binding of the GTP-binding protein to R* also lead to the formation of M II. At 2 degrees C, pH 8.3, in particular where no M II is formed in the absence of GTP-binding protein, binding signals and light-induced attachment of the GTP-binding protein to the membrane are still observed. Consistently, addition of GTP-binding protein to a suspension of extracted membranes bleached at 2 degrees C (pH 8.3) shifts the metarhodopsin I in equilibrium metarhodopsin II equilibrium towards metarhodopsin II. The shift is reversed by GTP, which dissociates the rhodopsin--GTP-binding protein complex. 4. At low ionic strength, where the GTP-binding protein is soluble in the dark (instead of being associated to the membrane as in the above experiments) M II still induces the binding whereas M I does not, indicating a much lower affinity of the GTP-binding protein for MI.
D000818:Animals; D001665:Binding Sites / Q000528:radiation effects; D001798:Blood Proteins / Q000378:metabolism / Q000528:radiation effects*; D002417:Cattle; D019204:GTP-Binding Proteins; D008027:Light; D011956:Receptors, Cell Surface / Q000378:metabolism / Q000528:radiation effects*; D012168:Retinal Pigments / Q000378:metabolism* / Q000528:radiation effects*; D012243:Rhodopsin / Q000031:analogs & derivatives / Q000378:metabolism* / Q000528:radiation effects*; D012542:Scattering, Radiation; D013053:Spectrophotometry
D001798:Blood Proteins; D011956:Receptors, Cell Surface; D012168:Retinal Pigments; C013110:metarhodopsins; D012243:Rhodopsin; D019204:GTP-Binding Proteins