cGMP phosphodiesterase in rod and cone outer segments of the retina

R. L. Hurwitz, A. H. Bunt-Milam, M. L. Chang, J. A. Beavo

Research output: Contribution to journalArticle

77 Scopus citations

Abstract

Immunochemical, chromatographic, and sodium dodecyl sulfate gel electrophoresis studies suggest that immunologically related but distinct cyclic GMP phosphodiesterases are present in rod and cone outer segments of the retina. Immunocytochemical studies demonstrated that one monoclonal antibody (ROS-1) recognized a determinant present in both rod and cone outer segments, while another monoclonal antibody (ROS-2) only recognized rod outer segments. At least two peaks of phosphodiesterase activity could be separated by high-performance anion-exchange chromatography of retinal extracts. Both peaks were recognized by ROS-1. None of the first peak and only 80% of the second broad peak of activity were recognized by ROS-2. High-performance liquid chromatography profiles from human fovea and several other types of cone-enriched retina showed that most of the activity was contained in the first peak, suggesting that this activity was derived from cone outer segments. Conversely, the phosphodiesterase in rod-enriched preparations migrated predominantly in the second peak. Sodium dodecyl sulfate-gel electrophoresis indicated that this first peak contained a single large immunoreactive polypeptide (α') that migrated with the same mobility as a phosphorylase b standard and was distinct from the more rapidly migrating large immunoreactive polypeptides (α and β) present in a broad second peak. The second peak could be further separated into a first part that contained a doublet of two immunoreactive polypeptides (α and β) that migrated faster than phosphorylase b and a later part that contained only the most rapidly migrating polypeptide (β). All of the peaks could be activated by histone or transducin:GTP, implying that all contained a small 11-kDa inhibitory subunit (γ) of the enzyme. Since the larger (α') and smaller (β) immunoreactive polypeptides could be completely separated from the α polypeptide and from each other, yet still retain the ability to be activated by histone or transducin, the data suggest that only a single species of polypeptide-inhibitor complex (e.g. α'γ, αγ, or βγ) was required for histone or transducin:GTP activation

Original languageEnglish (US)
Pages (from-to)568-573
Number of pages6
JournalJournal of Biological Chemistry
Volume260
Issue number1
StatePublished - 1985

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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