A Semi-throughput Procedure for Assaying Plant NADP-malate Dehydrogenase Activity Using a Plate Reader.
Enzyme activity assay
NADP-MDH
NADP-malate dehydrogenase
R script
Spectrophotometry
TRX
Thioredoxin
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
20 Aug 2023
20 Aug 2023
Historique:
received:
08
03
2023
revised:
04
04
2023
accepted:
04
06
2023
medline:
28
8
2023
pubmed:
28
8
2023
entrez:
28
8
2023
Statut:
epublish
Résumé
Chloroplast NADP-dependent malate dehydrogenase (NADP-MDH) is a redox regulated enzyme playing an important role in plant redox homeostasis. Leaf NADP-MDH activation level is considered a proxy for the chloroplast redox status. NADP-MDH enzyme activity is commonly assayed spectrophotometrically by following oxaloacetate-dependent NADPH oxidation at 340 nm. We have developed a plate-adapted protocol to monitor NADP-MDH activity allowing faster data production and lower reagent consumption compared to the classic cuvette format of a spectrophotometer. We provide a detailed procedure to assay NADP-MDH activity and measure the enzyme activation state in purified protein preparations or in leaf extracts. This protocol is provided together with a semi-automatized data analysis procedure using an R script.
Identifiants
pubmed: 37638298
doi: 10.21769/BioProtoc.4769
pii: e4769
pmc: PMC10450749
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4769Informations de copyright
©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no conflict of interest.
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