Characterization of a phosphodiesterase-immunoreactive polypeptide from rod photoreceptors of developing rd mouse retinas.
In the inherited retinal degeneration of rd mice, cyclic GMP accumulates in affected rod photoreceptors prior to their degeneration. A deficiency in the activity of the visual cell phosphodiesterase apparently results in the accumulation of cyclic GMP. The cyclic GMP phosphodiesterase (PDE) of normal mouse photoreceptors is a heteromeric protein complex of about 170 kDa, consisting of the alpha beta catalytic unit and the gamma inhibitory unit. The isolated complex has low enzyme activity but it can be activated by incubation with histone. Affinity-purified polyclonal antibodies against the PDE complex of bovine rod outer segments were prepared and used to identify in retinas of both normal and rd mice PDE-immunoreactive polypeptides which comigrated on SDS-polyacrylamide gels with the large subunits (88 kDa) of the normal PDE complex. During development of normal retinas, the 88 kDa immunoreactive component of the PDE complex were detected by day 7, with immunoreactivity increasing throughout the second postnatal week. In rd retinas, the 88 kDa immunoreactivity increased after 9 postnatal days, decreased during rod photoreceptor degeneration, and was undetectable in mature rd retinas. Under nondenaturing conditions, the PDE-immunoreactive polypeptide of rd retinas sedimented on sucrose gradients with a sedimentation coefficient of 5.6S and an apparent molecular mass of about 105 kDa; no associated histone-activated PDE activity was detected. These findings show that PDE-immunoreactive polypeptides are synthesized in immature rd photoreceptors and that the PDE-immunoreactive polypeptides fail to form a PDE complex which is comparable to that of normal photoreceptors.
Lee
R H
RH
Department of Anatomy, University of California, Los Angeles.
Navon
S E
SE
Brown
B M
BM
Fung
B K
BK
Lolley
R N
RN
D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D002499:Centrifugation, Density Gradient; D007158:Immunologic Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains / Q000235:genetics / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D010727:Phosphoric Diester Hydrolases / Q000276:immunology*; D010786:Photoreceptor Cells / Q000254:growth & development / Q000276:immunology*; D011817:Rabbits; D012016:Reference Values; D012160:Retina / Q000254:growth & development* / Q000276:immunology; D012162:Retinal Degeneration / Q000235:genetics* / Q000276:immunology / Q000503:physiopathology
D010455:Peptides; D010727:Phosphoric Diester Hydrolases