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Redox cycling and kinetic analysis of single molecules of solution-phase nitrite reductase

Lookup NU author(s): Dorota Kostrz, Professor Christopher DennisonORCiD

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Abstract

Single-molecule measurements are a valuable tool for revealing details of enzyme mechanisms by enabling observation of unsynchronized behavior. However, this approach often requires immobilizing the enzyme on a substrate, a process which may alter enzyme behavior. We apply a microfluidic trapping device to allow, for the first time, prolonged solution-phase measurement of single enzymes in solution. Individual redox events are observed for single molecules of a blue nitrite reductase and are used to extract the microscopic kinetic parameters of the proposed catalytic cycle. Changes in parameters as a function of substrate concentration are consistent with a random sequential substrate binding mechanism.

Publication metadata

Author(s): Goldsmith RH, Tabares LC, Kostrz D, Dennison C, Aartsma TJ, Canters GW, Moerner WE

Publication type: Article

Publication status: Published

Journal: Proceedings of the National Academy of Sciences

Year: 2011

Volume: 108

Issue: 42

Pages: 17269-17274

Print publication date: 18/10/2011

ISSN (print): 0027-8424

ISSN (electronic): 1091-6490

Publisher: National Academy of Sciences

URL: http://dx.doi.org/10.1073/pnas.1113572108

DOI: 10.1073/pnas.1113572108

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