✦ LIBER ✦

A continuous fluorescence assay for cyclic nucleotide phosphodiesterase hydrolysis of cyclic GMP

✍ Scribed by J.David Johnson; John D. Walters; John S. Mills

Publisher: Elsevier Science
Year: 1987
Tongue: English
Weight: 376 KB
Volume: 162
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(87)90039-x

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✦ Synopsis

The fluorescent 2'-methylanthraniloyl derivative of cyclic GMP undergoes a 45% decrease in fluorescence when it is cleaved by brain phosphodiesterase in the presence of calmodulin. This fluorescence decrease is dependent upon calcium, calmodulin, and phosphodiesterase, and correlates well (r = 0.996) with the disappearance of substrate as monitored by high-performance liquid chromatography. The Kd values determined by this fluorescence method and HPLC suggest that cyclic GMP and its fluorescent derivative exhibit similar kinetic parameters in their hydrolysis.

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