The flavoenzyme ferredoxin-NADP+ reductase (FNR) catalyzes the production of NADPH during photosynthesis. Whereas the structures of FNRs from spinach leaf and a cyanobacterium as well as many of their homologs have been solved, none of these studies has yielded a productive geometry of the flavin-nicotinamide interaction. Here, we show that this failure occurs because nicotinamide binding to wild type FNR involves the energetically unfavorable displacement of the C-terminal Tyr side chain. We used mutants of this residue (Tyr 308) of pea FNR to obtain the structures of productive NADP+ and NADPH complexes. These structures reveal a unique NADP+ binding mode in which the nicotinamide ring is not parallel to the flavin isoalloxazine ring, but lies against it at an angle of approximately 30 degrees, with the C4 atom 3 A from the flavin N5 atom.

A productive NADP+ binding mode of ferredoxin-NADP+ reductase revealed by protein engineering and crystallographic studies / Z. Deng, A. Aliverti, G. Zanetti, A.K. Arakaki, J. Ottado, E.G. Orellano, N.B. Calcaterra, E.A. Ceccarelli, N. Carrillo, P.A. Karplus. - In: NATURE STRUCTURAL BIOLOGY. - ISSN 1072-8368. - 6:9(1999 Sep), pp. 847-853. [10.1038/12307]

A productive NADP+ binding mode of ferredoxin-NADP+ reductase revealed by protein engineering and crystallographic studies

A. Aliverti
Secondo
;
G. Zanetti;
1999-09

Abstract

The flavoenzyme ferredoxin-NADP+ reductase (FNR) catalyzes the production of NADPH during photosynthesis. Whereas the structures of FNRs from spinach leaf and a cyanobacterium as well as many of their homologs have been solved, none of these studies has yielded a productive geometry of the flavin-nicotinamide interaction. Here, we show that this failure occurs because nicotinamide binding to wild type FNR involves the energetically unfavorable displacement of the C-terminal Tyr side chain. We used mutants of this residue (Tyr 308) of pea FNR to obtain the structures of productive NADP+ and NADPH complexes. These structures reveal a unique NADP+ binding mode in which the nicotinamide ring is not parallel to the flavin isoalloxazine ring, but lies against it at an angle of approximately 30 degrees, with the C4 atom 3 A from the flavin N5 atom.
photosynthesis ; protein ; enzyme ; hydride transfer ; biological oxidoreduction ; charge-transfer complex ; protein engineering ; flavin nucleotide ; nicotinamide dinucleotide
Settore BIO/10 - Biochimica
Settore BIO/11 - Biologia Molecolare
Settore BIO/04 - Fisiologia Vegetale
http://www.nature.com/nsmb/journal/v6/n9/full/nsb0999_847.html
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/189875
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