The regulation of the cGMP-binding cGMP phosphodiesterase by proteins that are immunologically related to γ subunit of the photoreceptor cGMP phosphodiesterase

Abstract

The cGMP phosphodiesterase from retinal rods (PDE-6) is an αβγ2 heterotetramer. The α and β subunits contain catalytic sites for cGMP hydrolysis, whereas the subunits serve as a protein inhibitor of the enzyme. Visual excitation of photoreceptors enables the activated GTP-bound form of the G-protein transducin to remove the inhibitory action of the γ subunit, thereby triggering PDE-6 activation. The type 5 phosphodiesterase (PDE-5) isoform shares a number of similar characteristics with PDE-6, including binding of cGMP to non catalytic sites, the cyclic nucleotide specificity, and inhibitor sensitivities. Although the functional role of PDE-5 remains unclear, it has been shown to be activated by protein kinase A (PKA) (Burns, F., Rodger, I.W. and Pyne, N.J. (1992) Biochem. J. 283, 487-491). Here we report that both the recombinant γ subunit and a peptide corresponding to amino acids 24-46 in this protein inhibited the activation of PDE-5 by PICA. Furthermore, immunoblotting airway smooth muscle membranes with a specific antibody against amino acids 24-46 of the PDE-6 γ subunit identified two major immunoreactive small molecular mass proteins of 14 and 18 kDa (p14 and p18). These appear to form a complex with PDE-5, be cause PDE activity was immunoprecipitated using antibody against the PDE-6 γ subunit. p14 and p18 were also substrates for phosphorylation by a unidentified kinase that was stimulated by a pertussis toxin-sensitive G protein. Phosphorylation of p14/p18 in membranes treated with guanine nucleotides correlated with a can current reduction in the activation of PDE-5 by PKA. We suggest that p14 and p18 share an epitope common to PDE-6 γ and that this region may interact with PDE-5 to prevent its activation by PKA.