The enzyme mechanism of nitrite reductase studied at single-molecule level

Kuznetsova, Sofya, Zauner, Gerhild, Aartsma, Thijs J., Engelkamp, Hans, Hatzakis, Nikos, Rowan, Alan E., Nolte, Roeland J. M., Christianen, Peter C. M. and Canters, Gerard W. (2008) The enzyme mechanism of nitrite reductase studied at single-molecule level. Proceedings of the National Academy of Sciences, 105 9: 3250-3255. doi:10.1073/pnas.0707736105

Author Kuznetsova, Sofya
Zauner, Gerhild
Aartsma, Thijs J.
Engelkamp, Hans
Hatzakis, Nikos
Rowan, Alan E.
Nolte, Roeland J. M.
Christianen, Peter C. M.
Canters, Gerard W.
Title The enzyme mechanism of nitrite reductase studied at single-molecule level
Journal name Proceedings of the National Academy of Sciences   Check publisher's open access policy
ISSN 0027-8424
Publication date 2008-03-04
Year available 2008
Sub-type Article (original research)
DOI 10.1073/pnas.0707736105
Open Access Status Not yet assessed
Volume 105
Issue 9
Start page 3250
End page 3255
Total pages 6
Place of publication Washington, DC, United States
Publisher National Academy of Sciences
Language eng
Abstract A generic method is described for the fluorescence “readout” of the activity of single redox enzyme molecules based on Förster resonance energy transfer from a fluorescent label to the enzyme cofactor. The method is applied to the study of copper-containing nitrite reductase from Alcaligenes faecalis S-6 immobilized on a glass surface. The parameters extracted from the single-molecule fluorescence time traces can be connected to and agree with the macroscopic ensemble averaged kinetic constants. The rates of the electron transfer from the type 1 to the type 2 center and back during turnover exhibit a distribution related to disorder in the catalytic site. The described approach opens the door to single-molecule mechanistic studies of a wide range of redox enzymes and the precise investigation of their internal workings.
Keyword Electron transfer
Fluorescent label
Förster transfer
Nitric oxide
Redox enzyme
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
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