Speeding up sequence specific assignment of IDPs.
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.
Bruker BioSpin GmbH, Silberstreifen, 76287 Rheinstetten, Germany.
Wolfgang
W
Bermel
Ivano
I
Bertini
Isabella C
IC
Felli
Leonardo
L
Gonnelli
Wiktor
W
Koźmiński
Alessandro
A
Piai
Roberta
R
Pierattelli
Jan
J
Stanek
D002247:Carbon Isotopes; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D051844:alpha-Synuclein / Q000737:chemistry
D002247:Carbon Isotopes; D011506:Proteins; D051844:alpha-Synuclein