Assay for phosphodiesterase function

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

Reexamination Certificate

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C435S007200, C435S007400, C435S007720, C435S007900, C435S008000, C435S015000, C435S019000, C435S021000, C435S196000

Reexamination Certificate

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10824771

ABSTRACT:
The present invention provides filter-based assays for measuring binding of compounds to phosphodiesterases (PDEs). The assay permits stoichiometric binding of compounds to PDEs, thereby providing a highly sensitive measure of a PDE binding and inhibition.

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patent: 2005/0164298 (2005-07-01), Golz et al.
Ballard et al., “Effects of sildenafil on the relaxation of human corpus cavernosum tissue in vitro and on the activities of cyclic nucleotied phosphodiesterase isozymes,”J. Urol., 159:2164-2171, 1998.
Corbin et al., “Phosphorylation of phosphodiesterase-5 by cyclic meucleotide-dependent protein kinase alters its catalytic and allosteric cGMP-binding activities,”Eur. J. Biochem., 267:2760-2767, 2000.
Corbin et al., “[3H]sildenafil binding to phosphodiesterase-5 is specific, kinetically heterogeneous, and stimulated by cGMP,”Mol. Pharmacol., 63(6):1364-1372, 2003.
Corbin and Francis, “Cyclic GMP phosphodiesterase-5: target of sildenafil,”J. Biol. Chem., 274(20):13729-13732, 1999.
Francis et al., “Phosphorylation of isolated human phosphodiesterase-5 regulatory domain induces an apparent conformational change and increases cGMP binding affinity,”J. Biol. Chem., 277(49):47581-47587, 2002.
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Gopal et al., “Allosteric sites of phosphodiesterase-5 (PDE5): a potential role in negative feedback regulationo f cGMP signaling in corpus cavernosum,”Eur. J. Biochem., 268:3304-3312, 2001.
Okada and Asakawa, “Allosteric activation of cGMP-specific, cGMP-binding phosphodiesterase (PDE5) by cGMP,”Biochemistry, 41:9672-9679, 2002.
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