Cyclic nucleotide phosphodiesterases in cultured normal and RCS rat pigment epithelium: kinetics of cyclic AMP and cyclic GMP hydrolysis.
Kinetically distinct classes of cyclic AMP (cAMP) and cyclic GMP (cGMP) phosphodiesterase activities (PDEs) were detected in homogenates of cultured pigment epithelium (PE) from both normal and Royal College of Surgeons (RCS) rats. PDE activities with apparent low Michaelis constants (Low Km cAMP- and cGMP-PDEs) were associated with the supernatant, while PDE activities with apparent high Michaelis constants (high Km cAMP- and cGMP-PDEs) were slightly higher in the pellet than in the supernatant after ultracentrifugation (100,000 g). Activity of the low Km PDEs was significantly reduced while that of high Km PDEs was not affected by known inhibitors of PDE. In both normal and RCS rat PE low Km PDEs required calcium and magnesium ions for optimal activity while the high Km PDEs required neither. In homogenates of cultured RCS rat pigment epithelium (PE), the kinetic parameters for cAMP- and cGMP-PDEs were comparable to normal, with the exception of the Km value of the low Km cGMP-PDE. This Km value was two-fold higher in the RCS compared with the normal (indicative of a reduced affinity for cGMP). It remains to be determined if the reduced affinity for cGMP in the RCS PE is related to the genetic defect which is expressed as a deficiency in the phagocytosis of outer segments by these cells.
Kurtz
M J
MJ
Department of Ophthalmology, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston, MA 02114.
Edwards
R B
RB
Schmidt
S Y
SY
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism*; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D004789:Enzyme Activation; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains
D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D006152:Cyclic GMP; D008274:Magnesium; D002118:Calcium