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Nucleotidyl cyclase activity of soluble guanylyl cyclase in intact cells.

Abstract：

:Soluble guanylyl cyclase (sGC) is activated by nitric oxide (NO) and generates the second messenger cyclic GMP (cGMP). Recently, purified sGC α1β1 has been shown to additionally generate the cyclic pyrimidine nucleotides cCMP and cUMP. However, since cyclic pyrimidine nucleotide formation occurred only the presence of Mn(2+) but not Mg(2+), the physiological relevance of these in vitro findings remained unclear. Therefore, we studied cyclic nucleotide formation in intact cells. We observed NO-dependent cCMP- and cUMP formation in intact HEK293 cells overexpressing sGC α1β1 and in RFL-6 rat fibroblasts endogenously expressing sGC, using HPLC-tandem mass spectrometry. The identity of cCMP and cUMP was unambiguously confirmed by HPLC-time-of-flight mass spectrometry. Our data indicate that cCMP and cUMP play second messenger roles and that Mn(2+) is a physiological sGC cofactor.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Bähre H,Danker KY,Stasch JP,Kaever V,Seifert R

doi

10.1016/j.bbrc.2013.12.108

subject

Has Abstract

pub_date

2014-01-24 00:00:00

pages

1195-9

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(13)02179-7

journal_volume

443

pub_type

杂志文章

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