Localization of S-nitrosothiols and assay of nitric oxide synthase and S-nitrosoglutathione reductase activity in plants.
The study of the metabolism of nitric oxide and other reactive nitrogen species (RNS) in plants has been the subject of intensive work in the last decade due to the relevance of these molecules in the physiology and biochemistry of plants. However, there are still many methodological limitations in the specific and accurate determination and localization of RNS. This chapter describes several biochemical and cellular methods demonstrated to be useful for this purpose in different plant tissues. These methods are the determination of L-arginine-dependent nitric oxide synthase and S-nitrosoglutathione reductase activities, as well as cellular localization by confocal laser-scanning microscopy of S-nitrosothiols, particularly S-nitrosoglutathione. These approaches can help advance the knowledge of the function of RNS in plant cells.
Corpas
Francisco J
FJ
Departamento de Bioquimica, Biología Celular y Molecular de plantas, Estación Experimental del Zaidín CSIC, Apartado 419, E-18080 Grannada, Spain. javier.corpas@eez.csic.es
Carreras
Alfonso
A
Esteban
Francisco J
FJ
Chaki
Mounira
M
Valderrama
Raquel
R
del Río
Luis A
LA
Barroso
Juan B
JB
D000445:Aldehyde Oxidoreductases / Q000032:analysis*; D000818:Animals; D001120:Arginine / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D007150:Immunohistochemistry / Q000379:methods; D008163:Luminescent Measurements / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D008954:Models, Biological; D019001:Nitric Oxide Synthase / Q000032:analysis*; D010126:Ozone / Q000737:chemistry; D018515:Plant Leaves / Q000378:metabolism; D010944:Plants / Q000201:enzymology / Q000378:metabolism*; D026403:S-Nitrosothiols / Q000032:analysis*; D013053:Spectrophotometry / Q000379:methods; D014018:Tissue Distribution
D026403:S-Nitrosothiols; D010126:Ozone; D001120:Arginine; D019001:Nitric Oxide Synthase; D000445:Aldehyde Oxidoreductases; C087006:formaldehyde dehydrogenase, glutathione-independent