Affinity labeling of spinach ferredoxin-NADP+ oxidoreductase with periodate-oxidized NADP+.

Abstract:

:Periodate-oxidized NADP+ (dialdehyde-NADP+) inactivated soluble ferredoxin-NADP+ oxidoreductase and combined covalently to the enzyme. This inactivation was first order with respect to dialdehyde-NADP+ and followed saturation kinetics, indicating that the enzyme initially forms a reversible complex with the inactivator. NADP+ afforded complete protection against inactivation, while spinach ferredoxin was uneffective. In the presence of exogenous ferredoxin and illuminated thylakoids, the nucleotide analog functioned as a coenzyme for the reductase, although with rather lower efficiency than NADP+. It also acted as a competitive inhibitor with respect to NADPH in diaphorase activity. Incorporation of radioactivity from periodate-oxidized [3H]NADP+ gave a stoichiometry of 0.85 mol of reagent/mol of reductase, indicating that the modification of a single residue in the flavoprotein is responsible for the loss of enzymatic activity.

journal_name

Arch Biochem Biophys

authors

Chan RL,Carrillo N

doi

10.1016/0003-9861(84)90160-7

subject

Has Abstract

pub_date

1984-02-15 00:00:00

pages

340-7

issue

1

eissn

0003-9861

issn

1096-0384

pii

0003-9861(84)90160-7

journal_volume

229

pub_type

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