The characterization of intrinsically disordered proteins (IDPs) by NMR spectroscopy is made difficult by the extensive spectral overlaps. To overcome the intrinsic low-resolution of the spectra the introduction of high-dimensionality experiments is essential. We present here a set of high-resolution experiments based on direct (13)C-detection which proved useful in the assignment of α-synuclein, a paradigmatic IDP. In particular, we describe the implementation of 4D HCBCACON, HCCCON, HCBCANCO, 4/5D HNCACON and HNCANCO and 3/4D HCANCACO experiments, specifically tailored for spin system identification and backbone resonances sequential assignment. The use of non-uniform-sampling in the indirect dimension and of the H-flip approach to achieve longitudinal relaxation enhancement rendered the experiments very practical.

Speeding up sequence specific assignment of IDPs / W. Bermel; I. Bertini; I.C. Felli; L. Gonnelli; W. Koźmiński; A. Piai; R. Pierattelli; J. Stanek. - In: JOURNAL OF BIOMOLECULAR NMR. - ISSN 0925-2738. - STAMPA. - 53:(2012), pp. 293-301. [10.1007/s10858-012-9639-0]

Speeding up sequence specific assignment of IDPs

BERTINI, IVANO;FELLI, ISABELLA CATERINA;GONNELLI, LEONARDO;PIAI, ALESSANDRO;PIERATTELLI, ROBERTA;
2012

Abstract

The characterization of intrinsically disordered proteins (IDPs) by NMR spectroscopy is made difficult by the extensive spectral overlaps. To overcome the intrinsic low-resolution of the spectra the introduction of high-dimensionality experiments is essential. We present here a set of high-resolution experiments based on direct (13)C-detection which proved useful in the assignment of α-synuclein, a paradigmatic IDP. In particular, we describe the implementation of 4D HCBCACON, HCCCON, HCBCANCO, 4/5D HNCACON and HNCANCO and 3/4D HCANCACO experiments, specifically tailored for spin system identification and backbone resonances sequential assignment. The use of non-uniform-sampling in the indirect dimension and of the H-flip approach to achieve longitudinal relaxation enhancement rendered the experiments very practical.
2012
53
293
301
W. Bermel; I. Bertini; I.C. Felli; L. Gonnelli; W. Koźmiński; A. Piai; R. Pierattelli; J. Stanek
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/656161
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