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Ligand binding and conformational dynamics in a flavin-based electron-bifurcating enzyme complex revealed by Hydrogen-Deuterium Exchange Mass Spectrometry.
- Source :
-
FEBS letters [FEBS Lett] 2016 Dec; Vol. 590 (24), pp. 4472-4479. Date of Electronic Publication: 2016 Dec 16. - Publication Year :
- 2016
-
Abstract
- Flavin-based electron bifurcation (FBEB) is a novel mechanism of energy coupling used by anaerobic microorganisms to optimize their energy metabolism efficiency. The first high-resolution structure of a complete FBEB enzyme complex, the NADH-dependent reduced ferredoxin: NADP <superscript>+</superscript> -oxidoreductase (NfnAB) of Thermotoga maritima, was recently solved. However, no experimental evidence for the NADPH-binding site and conformational changes during the FBEB reaction are available. Here we analyzed ligand binding and the conformational dynamics of oxygen-sensitive NfnAB using Hydrogen-Deuterium Exchange Mass-Spectrometry, including a customized anaerobic workflow. We confirmed the NADH and the previously postulated NADPH-binding site. Furthermore, we observed an NfnA-NfnB rearrangement upon NADPH binding which supports the proposed FBEB mechanism.<br /> (© 2016 Federation of European Biochemical Societies.)
- Subjects :
- Amino Acid Sequence
Bacterial Proteins genetics
Bacterial Proteins metabolism
Binding Sites
Deuterium Exchange Measurement
Ferredoxins metabolism
Gene Expression
Ligands
Mass Spectrometry instrumentation
Mass Spectrometry methods
Models, Molecular
NAD metabolism
NADH, NADPH Oxidoreductases genetics
NADH, NADPH Oxidoreductases metabolism
NADP metabolism
Oxidation-Reduction
Protein Binding
Protein Domains
Protein Structure, Secondary
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Sequence Alignment
Substrate Specificity
Thermotoga maritima enzymology
Bacterial Proteins chemistry
Ferredoxins chemistry
NAD chemistry
NADH, NADPH Oxidoreductases chemistry
NADP chemistry
Thermotoga maritima chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1873-3468
- Volume :
- 590
- Issue :
- 24
- Database :
- MEDLINE
- Journal :
- FEBS letters
- Publication Type :
- Editorial & Opinion
- Accession number :
- 27889905
- Full Text :
- https://doi.org/10.1002/1873-3468.12489