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Superoxide dismutase and catalase are required to detect •NO from both coupled and uncoupled neuronal no synthase

Record type: Article

Despite numerous approaches to measuring nitric oxide (NO) formation from purified NO synthase (NOS), it is still not clear whether (.-)NO is a direct or indirect product of the NO synthase reaction. The direct detection of catalytically formed (.-)NO is complicated by side reactions with reactive oxide species like H(2)O(2) and superoxide. The aim of the present study was therefore to reinvestigate these reactions both electrochemically and by chemiluminescence detection with particular emphasis on the requirement for cofactors and their interference with (.-)NO detection. Flavins were found to generate large amounts of H(2)O(2) and were therefore excluded from subsequent incubations. Under conditions of both coupled and uncoupled catalysis, SOD was absolutely required to detect (.-)NO from NOS. H(2)O(2) formation took place also in the presence of SOD and gave a smaller yet significant interfering signal. Similar data were obtained when the proposed intermediate N(omega)-hydroxy-l-arginine was utilized as substrate. In conclusion, standard Clark-type ()NO electrodes are cross-sensitive to H(2)O(2) and therefore both SOD and catalase are absolutely required to specifically detect (.-)NO from NOS.

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Citation

Reif, A., Shutenko, Z. V., Feelisch, M. and Schmidt, H. H. H. W. (2004) Superoxide dismutase and catalase are required to detect •NO from both coupled and uncoupled neuronal no synthase Advances in Free Radical Biology & Medicine, 37, (7), pp. 988-997. (doi:10.1016/j.freeradbiomed.2004.07.005). (PMID:15336315).

More information

e-pub ahead of print date: 22 July 2004
Published date: 1 October 2004
Keywords: nitric oxide, NOS, superoxide, catalase, SOD, flavins, NO electrode, free radicals
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 337843
URI: http://eprints.soton.ac.uk/id/eprint/337843
ISSN: 8755-9668
PURE UUID: f7cfdf60-da2f-49c5-8442-6ca01e33d526
ORCID for M. Feelisch: ORCID iD orcid.org/0000-0003-2320-1158

Catalogue record

Date deposited: 07 Jun 2012 15:28
Last modified: 18 Jul 2017 06:00

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Contributors

Author: A. Reif
Author: Z. V. Shutenko
Author: M. Feelisch ORCID iD
Author: H. H. H. W. Schmidt

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