7Z0W

E. coli NfsA bound to NADP+

Summary for 7Z0W

• Entry DOI: 10.2210/pdb7z0w/pdb
• Descriptor: Oxygen-insensitive NADPH nitroreductase, FLAVIN MONONUCLEOTIDE, 2'-MONOPHOSPHOADENOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• Functional Keywords: oxygen-insensitive nadph nitroreductase, nadp+ complex, oxidoreductase
• Biological source: Escherichia coli K-12
• Total number of polymer chains: 8
• Total formula weight: 219375.04

Authors

White, S.A.,Grainger, A.,Parr, R.,Day, M.A.,Jarrom, D.,Graziano, A.,Searle, P.F.,Hyde, E.I. (deposition date: 2022-02-23, release date: 2022-07-20, Last modification date: 2024-01-31)

Primary citation

White, S.A.,Christofferson, A.J.,Grainger, A.I.,Day, M.A.,Jarrom, D.,Graziano, A.E.,Searle, P.F.,Hyde, E.I.
The 3D-structure, kinetics and dynamics of the E. coli nitroreductase NfsA with NADP + provide glimpses of its catalytic mechanism.
Febs Lett., 596:2425-2440, 2022

PubMed Abstract: Nitroreductases activate nitroaromatic antibiotics and cancer prodrugs to cytotoxic hydroxylamines and reduce quinones to quinols. Using steady-state and stopped-flow kinetics, we show that the Escherichia coli nitroreductase NfsA is 20-50 fold more active with NADPH than with NADH and that product release may be rate-limiting. The crystal structure of NfsA with NADP shows that a mobile loop forms a phosphate-binding pocket. The nicotinamide ring and nicotinamide ribose are mobile, as confirmed in molecular dynamics (MD) simulations. We present a model of NADPH bound to NfsA. Only one NADP is seen bound to the NfsA dimers, and MD simulations show that binding of a second NADP(H) cofactor is unfavourable, suggesting that NfsA and other members of this protein superfamily may have a half-of-sites mechanism.

PubMed: 35648111
DOI: 10.1002/1873-3468.14413

Experimental method

X-RAY DIFFRACTION (2.06 Å)