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Issue 5, 2013
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Characterisation of an intrinsically disordered protein complex of Swi5–Sfr1 by ion mobility mass spectrometry and small-angle X-ray scattering

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Abstract

It is now recognized that intrinsically disordered proteins (IDPs) play important roles as hubs in intracellular networks, and their structural characterisation is of significance. However, due to their highly dynamic features, it is challenging to investigate the structures of IDPs solely by conventional methods. In the present study, we demonstrate a novel method to characterise protein complexes using electrospray ionization ion mobility mass spectrometry (ESI-IM-MS) in combination with small-angle X-ray scattering (SAXS). This method enables structural characterisation even of proteins that have difficulties in crystallisation. With this method, we have characterised the Schizosaccharomyces pombe Swi5–Sfr1 complex, which is expected to have a long disordered region at the N-terminal portion of Sfr1. ESI-IM-MS analysis of the Swi5–Sfr1 complex revealed that its experimental collision cross-section (CCS) had a wide distribution, and the CCS values of the most dominant ions were ∼56% of the theoretically calculated value based on the SAXS low-resolution model, suggesting a significant size reduction in the gas phase. The present study demonstrates that the newly developed method for calculation of the theoretical CCSs of the SAXS low-resolution models of proteins allows accurate evaluation of the experimental CCS values of IDPs provided by ESI-IM-MS by comparing with the low-resolution solution structures. Furthermore, it was revealed that the combination of ESI-IM-MS and SAXS is a promising method for structural characterisation of protein complexes that are unable to crystallise.

Graphical abstract: Characterisation of an intrinsically disordered protein complex of Swi5–Sfr1 by ion mobility mass spectrometry and small-angle X-ray scattering

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Publication details

The article was received on 28 Jun 2012, accepted on 17 Dec 2012 and first published on 18 Dec 2012


Article type: Paper
DOI: 10.1039/C2AN35878F
Analyst, 2013,138, 1441-1449

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    Characterisation of an intrinsically disordered protein complex of Swi5–Sfr1 by ion mobility mass spectrometry and small-angle X-ray scattering

    K. Saikusa, N. Kuwabara, Y. Kokabu, Y. Inoue, M. Sato, H. Iwasaki, T. Shimizu, M. Ikeguchi and S. Akashi, Analyst, 2013, 138, 1441
    DOI: 10.1039/C2AN35878F

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