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Fast hydrogen exchange affects 15N relaxation measurements in intrinsically disordered proteins
Ist Teil von
Journal of biomolecular NMR, 2013-03, Vol.55 (3), p.249-256
Ort / Verlag
Dordrecht: Springer Netherlands
Erscheinungsjahr
2013
Link zum Volltext
Quelle
Alma/SFX Local Collection
Beschreibungen/Notizen
Unprotected amide protons can undergo fast hydrogen exchange (HX) with protons from the solvent. Generally, NMR experiments using the out-and-back coherence transfer with amide proton detection are affected by fast HX and result in reduced signal intensity. When one of these experiments,
1
H–
15
N HSQC, is used to measure the
15
N transverse relaxation rate (R
2
), the measured R
2
rate is convoluted with the HX rate (k
HX
) and has higher apparent R
2
values. Since the
15
N R
2
measurement is important for analyzing protein backbone dynamics, the HX effect on the R
2
measurement is investigated and described here by multi-exponential signal decay. We demonstrate these effects by performing
15
N R
2
CPMG
experiments on α-synuclein, an intrinsically disordered protein, in which the amide protons are exposed to solvent. We show that the HX effect on R
2
CPMG
can be extracted by the derived equation. In conclusion, the HX effect may be pulse sequence specific and results from various sources including the J coupling evolution, the change of steady state water proton magnetization, and the D
2
O content in the sample. To avoid the HX effect on the analysis of relaxation data of unprotected amides, it is suggested that NMR experimental conditions insensitive to the HX should be considered or that intrinsic R
2
CPMG
values be obtained by methods described herein.