Catalytic mechanism of hydride transfer between NADP(+)/H and ferredoxin-NADP(+) reductase from Anabaena PCC 7119

Jesus Tejero, Jose Ramon Peregrina, Marta Martinez-Julvez, Aldo Gutierrez, Carlos Gomez-Moreno, Nigel S. Scrutton, Milagros Medina

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41 Citations (Scopus)

Abstract

The mechanism of hydride transfer between Anabaena FNR and NADP(+)/H was analysed using for the first time stopped-flow photodiode array detection and global analysis deconvolution. The results indicated that the initial spectral changes, occurring within the instrumental dead time upon reaction of FNR with NADP(+)/H, included not only the initial interaction and complex formation, but also the first subsequent steps of the sequential reactions that involve hydride transfer. Two different charge-transfer complexes formed prior and upon hydride transfer, FNRox-NADPH and FNRrd-NADP(+). Detectable amounts of FNROox-NADPH were found at equilibrium, but FNRrd-NADP(+) accumulated to a small extent and quickly evolved. The spectral properties of both charge-transfer complexes, for the first time in Anabaena FNR, as well as the corresponding inter-conversion hydride transfer rates were obtained. The need of an adequate initial interaction between NADP(+)/H and FNR, and subsequent conformational changes, was also established by studying the reactions of two FNR mutants. (c) 2006 Elsevier Inc. All rights reserved.

Original languageEnglish
Pages (from-to)79-90
Number of pages12
JournalArchives of Biochemistry and Biophysics
Volume459
Issue number1
DOIs
Publication statusPublished - 1 Mar 2007

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