A soluble form of bovine rod photoreceptor phosphodiesterase has a novel 15-kDa subunit

P. G. Gillespie, R. K. Prusti, E. D. Apel, J. A. Beavo

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Abstract

A substantial fraction (20-30%) of the bovine rod outer segment phosphodiesterase (PDE) activity is not associated with outer segment membranes prepared with buffers of moderate ionic strength; this PDE activity appears to represent a distinct, soluble isozyme. Although this PDE isozyme can be demonstrated to be present in sealed rod outer segments, it is discarded from most standard rod outer segment preparations. A method was developed that allowed the rapid purification of the soluble rod PDE by 2600-fold, to apparent homogeneity, using a monoclonal antibody column (ROS-1a). The soluble rod PDE isozyme has a novel M(r) = 15,000 subunit (δ) in addition to subunits of M(r) = 88,000 (α(sol)), 84,000 (β(sol)), and 11,000 (γ(sol)). The δ subunit comigrates with and may be identical to the cone PDE 15-kDa subunit. The small subunits of the soluble rod PDE and the membrane-associated rod PDE were isolated by reverse-phase chromatography. The γ(sol) subunit was a potent inhibitor of trypsin-activated rod PDE, inhibiting 50% of 1 pM PDE activity at a concentration of 11 pM. This concentration was similar to that observed for the γ subunit of the membrane-associated rod PDE. The purified δ subunit did not appear to affect PDE activity; this subunit was, however, unusually difficult to keep in solution. All of the kinetic and physical properties of the soluble rod PDE tested thus far are similar to those of the membrane-associated form, except for the presence of the δ subunit, suggesting that this unique subunit could mediate the solubility of the soluble rod PDE and the cone PDE in the intact photoreceptor.

Original languageEnglish (US)
Pages (from-to)12187-12193
Number of pages7
JournalJournal of Biological Chemistry
Volume264
Issue number21
StatePublished - Jan 1 1989

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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