5VOH

Crystal structure of engineered water-forming NADPH oxidase (TPNOX) bound to NADPH. The G159A, D177A, A178R, M179S, P184R mutant of LbNOX.

Summary for 5VOH

• Entry DOI: 10.2210/pdb5voh/pdb
• Descriptor: NADH oxidase, FLAVIN-ADENINE DINUCLEOTIDE, NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, ... (7 entities in total)
• Functional Keywords: nadph, redox active cysteine, water-forming oxidase, oxidoreductase
• Biological source: Lactobacillus brevis KB290
• Total number of polymer chains: 4
• Total formula weight: 231654.66

Authors

Cracan, V.,Grabarek, Z. (deposition date: 2017-05-02, release date: 2017-08-02, Last modification date: 2024-10-09)

Primary citation

Cracan, V.,Titov, D.V.,Shen, H.,Grabarek, Z.,Mootha, V.K.
A genetically encoded tool for manipulation of NADP(+)/NADPH in living cells.
Nat. Chem. Biol., 13:1088-1095, 2017

PubMed Abstract: The redox coenzymes NADH and NADPH are broadly required for energy metabolism, biosynthesis and detoxification. Despite detailed knowledge of specific enzymes and pathways that utilize these coenzymes, a holistic understanding of the regulation and compartmentalization of NADH- and NADPH-dependent pathways is lacking, partly because of a lack of tools with which to investigate these processes in living cells. We have previously reported the use of the naturally occurring Lactobacillus brevis HO-forming NADH oxidase (LbNOX) as a genetic tool for manipulation of the NAD/NADH ratio in human cells. Here, we present triphosphopyridine nucleotide oxidase (TPNOX), a rationally designed and engineered mutant of LbNOX that is strictly specific to NADPH. We characterized the effects of TPNOX expression on cellular metabolism and used it in combination with LbNOX to show how the redox states of mitochondrial NADPH and NADH pools are connected.

PubMed: 28805804
DOI: 10.1038/nchembio.2454

Experimental method

X-RAY DIFFRACTION (2.302 Å)