A1 Journal article (refereed)
HACANCOi : a new Hα-detected experiment for backbone resonance assignment of intrinsically disordered proteins (2020)

Karjalainen, M., Tossavainen, H., Hellman, M., & Permi, P. (2020). HACANCOi : a new Hα-detected experiment for backbone resonance assignment of intrinsically disordered proteins. Journal of Biomolecular NMR, 74(12), 741-752. https://doi.org/10.1007/s10858-020-00347-5

JYU authors or editors

Publication details

All authors or editors: Karjalainen, Mikael; Tossavainen, Helena; Hellman, Maarit; Permi, Perttu

Journal or series: Journal of Biomolecular NMR

ISSN: 0925-2738

eISSN: 1573-5001

Publication year: 2020

Publication date: 28/10/2020

Volume: 74

Issue number: 12

Pages range: 741-752

Publisher: Springer

Publication country: Netherlands

Publication language: English

DOI: https://doi.org/10.1007/s10858-020-00347-5

Publication open access: Openly available

Publication channel open access: Partially open access channel

Publication is parallel published (JYX): https://jyx.jyu.fi/handle/123456789/72423

Abstract

Unidirectional coherence transfer is highly efficient in intrinsically disordered proteins (IDPs). Their elevated ps-ns timescale dynamics ensures long transverse (T2) relaxation times allowing sophisticated coherence transfer pathway selection in comparison to folded proteins. 1Hα-detection ensures non-susceptibility to chemical exchange with the solvent and enables chemical shift assignment of consecutive proline residues, typically abundant in IDPs. However, many IDPs undergo a disorder-to-order transition upon interaction with their target protein, which leads to the loss of the favorable relaxation properties. Long coherence transfer routes now result in prohibitively large decrease in sensitivity. We introduce a novel 4D 1Hα-detected experiment HACANCOi, together with its 3D implementation, which warrant high sensitivity for the assignment of proline-rich regions in IDPs in complex with a globular protein. The experiment correlates 1Hαi, 13Cαi, 15Ni and 13C′i13Ci′ spins by transferring the magnetization concomitantly from 13Cαi to 15Ni and 13C′i13Ci′. The B1 domain of protein G (GB1), and the enteropathogenic E. coli EspF in complex with human SNX9 SH3, serve as model systems to demonstrate the attainable sensitivity and successful sequential assignment.

Keywords: proteins; NMR spectroscopy

Free keywords: E. coli; EspF; GB1; intrinsically disordered protein; IDP; resonance assignment; SNX9 SH3

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Ministry reporting: Yes

Reporting Year: 2020

JUFO rating: 1