Exquisite sensitivity to subsecond, picomolar nitric oxide transients conferred on cells by guanylyl cyclase-coupled receptors

Batchelor, A.M., Bartus, K., Reynell, C., Constantinou, S., Halvey, E.J., Held, K.F., Dostmann, W.R., Vernon, J. and Garthwaite, J. 2010. Exquisite sensitivity to subsecond, picomolar nitric oxide transients conferred on cells by guanylyl cyclase-coupled receptors. Proceedings of the National Academy of Sciences of the United States of America. 107 (51), pp. 22060-22065.

TitleExquisite sensitivity to subsecond, picomolar nitric oxide transients conferred on cells by guanylyl cyclase-coupled receptors
AuthorsBatchelor, A.M., Bartus, K., Reynell, C., Constantinou, S., Halvey, E.J., Held, K.F., Dostmann, W.R., Vernon, J. and Garthwaite, J.
Abstract

Nitric oxide (NO) functions as a diffusible transmitter in most tissues of the body and exerts its effects by binding to receptors harboring a guanylyl cyclase transduction domain, resulting in cGMP accumulation in target cells. Despite its widespread importance, very little is known about how this signaling pathway operates at physiological NO concentrations and in real time. To address these deficiencies, we have exploited the properties of a novel cGMP biosensor, named δ-FlincG, expressed in cells containing varying mixtures of NO-activated guanylyl cyclase and cGMP-hydrolyzing phosphodiesterase activity. Responsiveness to NO, signifying a physiologically relevant rise in cGMP to 30 nM or more, was seen at concentrations as low as 1 pM, making cells by far the most sensitive NO detectors yet encountered. Even cells coexpressing phosphodiesterase-5, a cGMP-activated isoform found in many NO target cells, responded to NO in concentrations as low as 10 pM. The dynamics of NO capture and signal transduction was revealed by administering timed puffs of NO from a local pipette. A puff lasting only 100 ms, giving a calculated peak intracellular NO concentration of 23 pM, was detectable. The results could be encapsulated in a quantitative model of cellular NO-cGMP signaling, which recapitulates the NO responsiveness reported previously from crude cGMP measurements on native cells, and which explains how NO is able to exert physiological effects at extremely low concentrations, when only a tiny proportion of its receptors would be occupied.

JournalProceedings of the National Academy of Sciences of the United States of America
Journal citation107 (51), pp. 22060-22065
ISSN0027-8424
Year2010
PublisherNational Academy of Sciences
Digital Object Identifier (DOI)doi:10.1073/pnas.1013147107
Publication dates
PublishedDec 2010

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