5VW2

NADPH soak of Y316S mutant of corn root ferredoxin:NADP+ reductase

Summary for 5VW2

• Entry DOI: 10.2210/pdb5vw2/pdb
• Related: 5VW3 5VW4 5VW5 5VW6 5VW7 5VW8 5VW9 5VWA 5VWB
• Descriptor: Ferredoxin--NADP reductase, MAGNESIUM ION, ACETATE ION, ... (6 entities in total)
• Functional Keywords: flavoenzyme, hydride transfer, photosynthesis, active site compression, transferase, oxidoreductase
• Biological source: Zea mays (Maize)
• Total number of polymer chains: 1
• Total formula weight: 36900.22

Authors

Kean, K.M.,Carpenter, R.A.,Hall, A.R.,Karplus, P.A. (deposition date: 2017-05-21, release date: 2017-08-23, Last modification date: 2024-10-23)

Primary citation

Kean, K.M.,Carpenter, R.A.,Pandini, V.,Zanetti, G.,Hall, A.R.,Faber, R.,Aliverti, A.,Karplus, P.A.
High-resolution studies of hydride transfer in the ferredoxin:NADP(+) reductase superfamily.
FEBS J., 284:3302-3319, 2017

PubMed Abstract: Ferredoxin: NADP reductase (FNR) is an FAD-containing enzyme best known for catalysing the transfer of electrons from ferredoxin (Fd) to NADP to make NADPH during photosynthesis. It is also the prototype for a broad enzyme superfamily, including the NADPH oxidases (NOXs) that all catalyse similar FAD-enabled electron transfers between NAD(P)H and one-electron carriers. Here, we define further mechanistic details of the NAD(P)H ⇌ FAD hydride-transfer step of the reaction based on spectroscopic studies and high-resolution (~ 1.5 Å) crystallographic views of the nicotinamide-flavin interaction in crystals of corn root FNR Tyr316Ser and Tyr316Ala variants soaked with either nicotinamide, NADP , or NADPH. The spectra obtained from FNR crystal complexes match those seen in solution and the complexes reveal active site packing interactions and patterns of covalent distortion of the FAD that imply significant active site compression that would favour catalysis. Furthermore, anisotropic B-factors show that the mobility of the C4 atom of the nicotinamide in the FNR:NADP complex has a directionality matching that expected for boat-like excursions of the nicotinamide ring thought to enhance hydride transfer. Arguments are made for the relevance of this binding mode to catalysis, and specific consideration is given to how the results extrapolate to provide insight to structure-function relations for the membrane-bound NOX enzymes for which little structural information has been available.

PubMed: 28783258
DOI: 10.1111/febs.14190

Experimental method

X-RAY DIFFRACTION (1.451 Å)