title: Elevated level of protein phosphatase 2A activity in retinas of rd mice.
abstract: Phosphorylation of rhodopsin is not detectable in vitro in the retina of the rd mouse. We investigated the enzymatic system responsible for this abnormality by measuring the levels of rhodopsin kinase and protein phosphatase 2A in normal (rd/+) and diseased (rd/rd) mouse retinas of several ages. For each enzyme, we developed micro assays that were suitable for measuring enzyme activity in one-half mouse retina. Our results indicate that rhodopsin kinase activity is identical in rd/+ and rd/rd retinas until post-natal day 11, and it decreases thereafter in the rd/rd retina, correlating with the loss of rod photoreceptors that occurs in this tissue. Protein phosphatase 2A has a constant level of activity in rd/+ retinas from ages 5 to 32 days but it is higher than normal in rd/rd retinas from post-natal days 5 to 10. It then decreases to levels that are comparable to those in rd/+ retina. Although the rd/rd extract contains the elevated protein phosphatase 2A activity, when rd/rd and rd/+ retinal extracts are each subjected to gel filtration, the elution profiles of protein phosphatase 2A activity appear to be quantitatively identical. This apparent loss of rd/rd phosphatase activity suggests a difference in the regulatory behavior of the enzyme in the normal and degenerative retinas. Thus, the failure to detect in vitro phosphorylation of rhodopsin in the rd/rd retina seems to result from the elevated level of protein phosphatase 2A activity which could more rapidly remove the phosphate from phosphorylated rhodopsin. Since protein phosphatase 2A is a ubiquitous enzyme with broad specificity, an elevation in its activity also could affect other protein phosphorylations.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville.
authors.forename: K
authors.initials: K
authors.lastname: Palczewski
authors.forename: D B
authors.initials: DB
authors.lastname: Farber
authors.forename: P A
authors.initials: PA
authors.lastname: Hargrave
mesh_terms: D000375:Aging / Q000378:metabolism; D000818:Animals; D002850:Chromatography, Gel; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D054648:Protein Phosphatase 2; D012160:Retina / Q000201:enzymology*; D012162:Retinal Degeneration / Q000201:enzymology*; D012243:Rhodopsin / Q000378:metabolism
chemical_list: D005136:Eye Proteins; D012243:Rhodopsin; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; C496762:Grk1 protein, mouse; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2