Native Mass Spectrometry of Iron-Sulfur Proteins

Crack, Jason C. and Le Brun, Nick E. (2021) Native Mass Spectrometry of Iron-Sulfur Proteins. In: Methods in Molecular Biology. Methods in Molecular Biology . UNSPECIFIED, pp. 231-258. ISBN 978-1-0716-1605-5_13

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Abstract

Iron-sulfur clusters constitute a large and widely distributed group of protein cofactors that play key roles in a wide range of metabolic processes. The inherent reactivity of iron-sulfur clusters toward small molecules, for example, O 2, NO, or free Fe, makes them ideal for sensing changes in the cellular environment. Nondenaturing, or native, MS is unique in its ability to preserve the noncovalent interactions of many (if not all) species, including stable intermediates, while providing accurate mass measurements in both thermodynamic and kinetic experimental regimes. Here, we provide practical guidance for the study of iron-sulfur proteins by native MS, illustrated by examples where it has been used to unambiguously determine the type of cluster coordinated to the protein framework. We also describe the use of time-resolved native MS to follow the kinetics of cluster conversion, allowing the elucidation of the precise series of molecular events for all species involved. Finally, we provide advice on a unique approach to a typical thermodynamic titration, uncovering early, quasi-stable, intermediates in the reaction of a cluster with nitric oxide, resulting in cluster nitrosylation.

Item Type: Book Section
Uncontrolled Keywords: cluster conversion,iron-sulfur clusters,isotopic labeling,mass spectrometry,native ms,nondenaturing mass spectrometry,time-resolved ms,[2fe-2s],[3fe-4s],[4fe-4s],molecular biology,genetics ,/dk/atira/pure/subjectarea/asjc/1300/1312
Faculty \ School: Faculty of Science > School of Chemistry
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Depositing User: LivePure Connector
Date Deposited: 30 Jul 2021 23:40
Last Modified: 25 Aug 2021 23:45
URI: https://ueaeprints.uea.ac.uk/id/eprint/80928
DOI: 10.1007/978-1-0716-1605-5_13

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