Light-induced formation of NO in endothelial cells by photoactivatable NADPH analogues targeting nitric-oxide synthase.

Authors:
Rahima Chennoufi
Rahima Chennoufi
Université Paris-Saclay
Nicolas Bogliotti
Nicolas Bogliotti
Laboratoire de Chimie Organique
France
Bertrand Cinquin
Bertrand Cinquin
University of California San Francisco
United States
Patrick Tauc
Patrick Tauc
Centre Universitaire
France
Jean-Luc Boucher
Jean-Luc Boucher
Université Paris Descartes
France

Biochim Biophys Acta Gen Subj 2019 Jun 13;1863(6):1127-1137. Epub 2019 Apr 13.

LBPA, CNRS UMR8113, IDA FR3242, ENS Paris-Saclay, Université Paris-Saclay, F-94235 Cachan, France. Electronic address:

Background: Nitric-oxide synthases (NOS) catalyze the formation of NO using NADPH as electron donor. We have recently designed and synthesized a new series of two-photon absorbing and photoactivatable NADPH analogues (NT). These compounds bear one or two carboxymethyl group(s) on the 2'- or/and 3'-position(s) of the ribose in the adenosine moiety, instead of a 2'-phosphate group, and differ by the nature of the electron donor in their photoactivatable chromophore (replacing the nicotinamide moiety). Here, we addressed the ability of NTs to photoinduce eNOS-dependent NO production in endothelial cells.

Methods: The cellular fate of NTs and their photoinduced effects were studied using multiphoton fluorescence imaging, cell viability assays and a BODIPY-derived NO probe for NO measurements. The eNOS dependence of photoinduced NO production was addressed using two NOS inhibitors (NS1 and L-NAME) targeting the reductase and the oxygenase domains, respectively.

Results: We found that, two compounds, those bearing a single carboxymethyl group on the 3'-position of the ribose, colocalize with the Golgi apparatus (the main intracellular location of eNOS) and display high intracellular two-photon brightness. Furthermore, a eNOS-dependent photooxidation was observed for these two compounds only, which is accompanied by a substantial intracellular NO production accounting for specific photocytotoxic effects.

Conclusions: We show for the first time that NT photoactivation efficiently triggers electron flow at the eNOS level and increases the basal production of NO by endothelial cells.

General Significance: Efficient photoactivatable NADPH analogues targeting NOS could have important implications for generating apoptosis in tumor cells or modulating NO-dependent physiological processes.

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Source
http://dx.doi.org/10.1016/j.bbagen.2019.04.004DOI Listing
June 2019
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