Reduced Light-dependent Phosphorylation of an Analog Visual Pigment Containing 9-Demethylretinal as Its Chromophore (*)

9-Demethyl rhodopsin (9dR), an analog of vertebrate rhodopsin, consists of opsin and a covalently attached chromophore of 11-cis 9-demethylretinal. Electrophysiological evidence that photoactivated 9dR (9dR*) undergoes abnormally slow deactivation in salamander rods (Corson, D. W., Cornwall, M. C., and Pepperberg, D. R.(1994) Visual Neurosci. 11, 91-98) raises the possibility that opsin phosphorylation, a reaction involved in visual pigment deactivation, operates abnormally on 9dR*. This possibility was tested by measuring the light-dependent phosphorylation of 9dR in preparations obtained from bovine rod outer segments. Outer segment membranes containing 9dR or regenerated rhodopsin were flash-illuminated in the presence of [-P]ATP and rhodopsin kinase, further incubated in darkness, and then analyzed for opsin-bound [P]Pi. [P]Pi incorporation by 9dR* increased with both incubation period and bleaching extent but, under all conditions tested, was less than that measured in rhodopsin controls. Results obtained with 30-s incubation periods indicated that the maximal initial rate of incorporation by 9dR* is about 25% of that by photoactivated rhodopsin. The results imply that the low incorporation of Pi by 9dR* results from a reduced rate of phosphorylation by rhodopsin kinase and are consistent with the prolonged lifetime of 9dR* determined electrophysiologically.

[1]  K. Palczewski,et al.  Rod outer segment retinol dehydrogenase: substrate specificity and role in phototransduction. , 1994, Biochemistry.

[2]  H. Hamm,et al.  Regulation of transducin GTPase activity in bovine rod outer segments. , 1994, The Journal of biological chemistry.

[3]  E. MacNichol,et al.  Relief of opsin desensitization and prolonged excitation of rod photoreceptors by 9-desmethylretinal. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[4]  K. Palczewski,et al.  Opsins with mutations at the site of chromophore attachment constitutively activate transducin but are not phosphorylated by rhodopsin kinase. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[5]  J. Benovic,et al.  Visual arrestin binding to rhodopsin. Intramolecular interaction between the basic N terminus and acidic C terminus of arrestin may regulate binding selectivity. , 1994, The Journal of biological chemistry.

[6]  M. Cornwall,et al.  Evidence for the prolonged photoactivated lifetime of an analogue visual pigment containing 11 -cis 9-desmethylretinal , 1994, Visual Neuroscience.

[7]  K. Hofmann,et al.  Interaction between photoactivated rhodopsin and its kinase: stability and kinetics of complex formation. , 1993, Biochemistry.

[8]  R. Crouch,et al.  Mass spectrometric identification of phosphorylation sites in bleached bovine rhodopsin. , 1993, Biochemistry.

[9]  C. Morrison.  Polyethylene glycol as a hydration agent in oriented membrane bilayer samples. , 1993, Biophysical journal.

[10]  T. Lamb,et al.  Amplification and kinetics of the activation steps in phototransduction. , 1993, Biochimica et biophysica acta.

[11]  J. Jin,et al.  Light-dependent delay in the falling phase of the retinal rod photoresponse , 1992, Visual Neuroscience.

[12]  D. Morrison,et al.  Depalmitylation with hydroxylamine alters the functional properties of rhodopsin. , 1991, The Journal of biological chemistry.

[13]  L. Stryer.  Visual excitation and recovery. , 1991, The Journal of biological chemistry.

[14]  K. Palczewski,et al.  Regulation of rhodopsin kinase by autophosphorylation. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[15]  M. Bownds,et al.  Light activation of one rhodopsin molecule causes the phosphorylation of hundreds of others. A reaction observed in electropermeabilized frog rod outer segments exposed to dim illumination. , 1990, The Journal of biological chemistry.

[16]  David R. Pepperberg,et al.  Effect of bleached rhodopsin on signal amplification in rod visual receptors , 1990, Nature.

[17]  R. Rando,et al.  Schiff-base deprotonation is mandatory for light-dependent rhodopsin phosphorylation. , 1989, The Biochemical journal.

[18]  R. Rando,et al.  Removal of the 9-methyl group of retinal inhibits signal transduction in the visual process. A Fourier transform infrared and biochemical investigation. , 1989, Biochemistry.

[19]  K. Hofmann,et al.  Kinetics, binding constant, and activation energy of the 48-kDa protein-rhodopsin complex by extra-metarhodopsin II. , 1989, Biochemistry.

[20]  K. Palczewski,et al.  Purification and characterization of rhodopsin kinase. , 1988, The Journal of biological chemistry.

[21]  K. Hofmann,et al.  Photic modulation of a highly sensitive, near-infrared light-scattering signal recorded from intact retinal photoreceptors. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[22]  K. Palczewski,et al.  Rhodopsin kinase: substrate specificity and factors that influence activity. , 1988, Biochemistry.

[23]  N. Bennett,et al.  Inactivation of photoexcited rhodopsin in retinal rods: the roles of rhodopsin kinase and 48-kDa protein (arrestin). , 1988, Biochemistry.

[24]  J. Miller,et al.  Amplification of phosphodiesterase activation is greatly reduced by rhodopsin phosphorylation. , 1986, Biochemistry.

[25]  S. W. Hall,et al.  Phosphodiesterase activation by photoexcited rhodopsin is quenched when rhodopsin is phosphorylated and binds the intrinsic 48-kDa protein of rod outer segments. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[26]  U Wilden,et al.  Light-dependent phosphorylation of rhodopsin: number of phosphorylation sites. , 1982, Biochemistry.

[27]  M. Bitensky,et al.  Reciprocal effects of an inhibitory factor on catalytic activity and noncatalytic cGMP binding sites of rod phosphodiesterase. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[28]  S. Goff,et al.  The spectral properties of some visual pigment analogs. , 1973, Experimental eye research.

[29]  W. Hubbell,et al.  Lipid requirements for Rhodopsin regenerability. , 1973, Biochemistry.

[30]  H Ripps,et al.  S-Potentials in the Skate Retina , 1971, The Journal of general physiology.

[31]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[32]  V. Balasubramaniyan,et al.  A new series of synthetic visual pigments from cattle opsin and homologs of retinal. , 1969, Journal of the American Chemical Society.

[33]  G. Wald,et al.  THE MOLAR EXTINCTION OF RHODOPSIN , 1953, The Journal of general physiology.

[34]  E A Dratz,et al.  The molecular mechanism of visual excitation and its relation to the structure and composition of the rod outer segment. , 1987, Annual review of physiology.

[35]  R. Paulsen,et al.  Activation of rhodopsin phosphorylation is triggered by the lumirhodopsin–metarhodopsin I transition , 1983, Nature.