High pressure liquid chromatography of cyclic nucleotide phosphodiesterase from purified human T-lymphocytes.
The cyclic nucleotide phosphodiesterase (EC 3.1.4.17) in extracts of purified human peripheral blood T-lymphocytes was examined by ion exchange high pressure liquid chromatography. Four peaks of activity were isolated. The first peak of activity selectively hydrolyzed cyclic GMP. The following 3 peaks of activity (Ia, IIa and IIIa) were selective for cyclic AMP. The selective low Km cyclic AMP-phosphodiesterase inhibitor, Ro 20-1724 (d,1-1,4-[3-butoxy-4-methoxybenzyl]-2-imidazolidinone), did not inhibit the activity in Ia whereas it did inhibit the activity in IIa and IIIa (IC50 = 17 microM). The authors conclude that ion exchange high pressure liquid chromatography described in this communication is a useful method for the isolation of different forms of cyclic nucleotide phosphodiesterase activity from human T-lymphocytes.
Robicsek
S A
SA
Department of Pharmacology and Therapeutics, University of South Florida, College of Medicine, Tampa 33612-4799.
Krzanowski
J J
JJ
Szentivanyi
A
A
Polson
J B
JB
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000302:isolation & purification / Q000378:metabolism; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000302:isolation & purification; D012368:4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone / Q000494:pharmacology; D000818:Animals; D002147:Calmodulin / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D051381:Rats; D013601:T-Lymphocytes / Q000201:enzymology*
D002147:Calmodulin; D012368:4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D015106:3',5'-Cyclic-GMP Phosphodiesterases