A Guide to Native Mass Spectrometry to determine complex interactomes of molecular machines. (25th March 2020)

• Record Type: Journal Article
• Authors: Puglisi, Rita; Boeri Erba, Elisabetta; Pastore, Annalisa
• Is Part Of: FEBS journal. Volume 287:Number 12(2020)
• Page Start: 2428
• Page End: 2439
• Publication Date: 2020-03-25
• Subjects: Isc operon; FeS cluster; native mass spectrometry -- proteins -- structural biology; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 572
• DOI: 10.1111/febs.15281
• Languages: English
• ISSNs: 1742-464X

Abstract

Abstract : Native mass spectrometry is an emerging technique in biology that gives the possibility to study noncovalently bound complexes with high sensitivity and accuracy. It thus allows the characterization of macromolecular assemblies, assessing their mass and stoichiometries and mapping the interacting surfaces. In this review, we discuss the application of native mass spectrometry to dynamic molecular machines based on multiple weak interactions. In the study of these machines, it is crucial to understand which and under which conditions various complexes form at any time point. We focus on the specific example of the iron–sulfur cluster biogenesis machine because this is an archetype of a dynamic machine that requires very specific and demanding experimental conditions, such as anaerobicity and the need of retaining the fold of marginally folded proteins. We describe the advantages, challenges and current limitations of the technique by providing examples from our own experience and suggesting possible future solutions. Abstract : Native mass spectrometry (MS) can successfully be applied to study the interactome of iron–sulfur cluster biogenesis. In this review, we discuss how native MS spectra can determine all the species present in solution, their stoichiometry and their interactions.