Knapman, TW, Valette, NM, Warriner, SL et al. (1 more author) (2013) Ion mobility spectrometry-mass spectrometry of intrinsically unfolded proteins: trying to put order into disorder. Current Analytical Chemistry, 9 (2). 181 - 191. ISSN 1573-4110
Abstract
Intrinsically disordered proteins do not adopt well-defined native structures and therefore present an intriguing challenge in terms of structural elucidation as they are relatively inaccessible to traditional approaches such as NMR and X-ray crystallography. Many members of this important group of proteins have a distinct biological function and frequently undergo a conformational change on binding to their physiological targets which can in turn modulate their function. Furthermore, many intrinsically unstructured proteins are associated with a wide range of major diseases including cancer and amyloid-related disorders. Here, electrospray ionisation-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS) has been used to probe the conformational characteristics of two intrinsically disordered proteins: apo-cytochrome c and apo-osteocalcin. Both proteins are structured in their holo-states when bound to their respective substrates, but disordered in their apo-states. Here, the conformational properties of the holo- and the apo-protein forms for both species have been analysed and their mass spectral data and ion mobility spectrometry-derived collision cross-sectional areas, indicative of their physical size, compared to study the relationship between substrate binding and tertiary structure. In both cases, the intrinsically unstructured apo-states populated multiple conformations with larger cross-sectional areas than their holo-analogues, suggesting that intrinsic disorder in proteins does not preclude the formation of preferred conformations. Additionally, analysis of truncated analogues of osteocalcin has located the region of the protein responsible for the conformational changes detected upon metal cation binding. Together, the data illustrate the scope and utility of ESI-IMS-MS for studying the characteristics and properties of intrinsically disordered proteins whose analysis by other techniques is limited.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2013, Bentham Science Publishers. This is an author produced version of a paper published in Current Analytical Chemistry. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Collision cross-sectional areas; cytochrome c; electrospray ionisation; intrinsically disordered proteins; ion mobility spectrometry; mass spectrometry; osteocalcin |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 17 Feb 2015 12:52 |
Last Modified: | 30 Jun 2020 14:49 |
Published Version: | http://dx.doi.org/10.2174/1573411011309020004 |
Status: | Published |
Publisher: | Bentham Science Publishers |
Identification Number: | 10.2174/1573411011309020004 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:83111 |