IMR Press / FBL / Volume 23 / Issue 11 / DOI: 10.2741/4694

Frontiers in Bioscience-Landmark (FBL) is published by IMR Press from Volume 26 Issue 5 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on imrpress.com as a courtesy and upon agreement with Frontiers in Bioscience.

Open Access Review

Exploring the conformations of nitric oxide synthase with fluorescence

Show Less
1 Fourth Department of Internal Medicine, Evangelismos General Hospital, Athens, Greece
Front. Biosci. (Landmark Ed) 2018, 23(11), 2133–2146; https://doi.org/10.2741/4694
Published: 1 June 2018
(This article belongs to the Special Issue Nitric oxide: Biology and chemistry)
Abstract

Multi-domain oxidoreductases are a family of enzymes that catalyze oxidation-reduction reactions through a series of electron transfers. Efficient electron transfer requires a sequence of protein conformations that position electron donor and acceptor domains in close proximity to each other so that electron transfer can occur efficiently. An example is mammalian nitric oxide synthase (NOS), which consists of an N-terminal oxygenase domain containing heme and a C-terminal reductase domain containing NADPH/FAD and FMN subdomains. We describe the use of time-resolved and single-molecule fluorescence to detect and characterize the conformations and conformational dynamics of the neuronal and endothelial isoforms of NOS. Fluorescence signals are provided by a fluorescent dye attached to the Ca2+-signaling protein calmodulin (CaM), which regulates NOS activity. Time-resolved fluorescence decays reveal the presence of at least four underlying conformational states that are differentiated by the extent of fluorescence quenching. Single-molecule fluorescence displays transitions between conformational states on the time scales of milliseconds to seconds. This review describes the type of information available by analysis of time-resolved and single-molecule fluorescence experiments.

Keywords
Nitric Oxide Synthase
Fluorescence
FRET
Single-Molecule
Conformational Dynamics
Review
Share
Back to top