Subspecies of arrestin from bovine retina. Equal functional binding to photoexcited rhodopsin but various isoelectric focusing phenotypes in individuals.
Arrestin (also named 48-kDa protein or S-antigen) binds to photoexcited and phosphorylated rhodopsin and thereby prevents activation of cGMP phosphodiesterase (EC 3.1.4.35) by transducin in retinal rods. We report here that retinal arrestin consists of several subspecies (isoelectric points between pH 5.5-6.2), which can be separated by FPLC anion-exchange chromatography and by FPLC chromatofocusing resulting in highly enriched individual subspecies. The entire heterogeneity pattern of arrestin is present in rod outer segments, independently of whether arrestin orginated from the outer or mostly from the inner segment of rod cells. The different subspecies show a similar binding behavior to photoexcited rhodopsin phosphorylated to various degrees and they quench the cGMP phosphodiesterase activity equally well. In the presence of rod outer segment membranes, arrestin is phosphorylated light-dependently by protein kinase C (0.2 mol phosphate/mol arrestin). This implies that the heterogeneity of arrestin is not primarily due to phosphorylation. Arrestin from different individuals exists as four isoelectric focusing patterns which occur with remarkably different frequencies in calf and cattle. The complexity of the IEF pattern does not increase with aging. Distinct subspecies of arrestin may reflect differences in their primary structure, or may result from differentially regulated post-translational modifications in individuals.
Weyand
I
I
Institut für Biologische Informationsverarbeitung, Forschungszentrum Jülich GmbH, Federal Republic of Germany.
Kühn
H
H
D000818:Animals; D000941:Antigens / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D019393:Arrestin; D002417:Cattle; D002852:Chromatography, Ion Exchange; D006152:Cyclic GMP; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D007525:Isoelectric Focusing; D010449:Peptide Mapping; D010641:Phenotype; D010726:Phosphodiesterase Inhibitors; D010766:Phosphorylation; D011153:Population; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism; D011817:Rabbits; D012160:Retina / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000201:enzymology / Q000378:metabolism
D000941:Antigens; D019393:Arrestin; D005136:Eye Proteins; D010726:Phosphodiesterase Inhibitors; D012243:Rhodopsin; D011493:Protein Kinase C; D006152:Cyclic GMP