Real-time pure shift ¹⁵N HSQC of proteins: a real improvement in resolution and sensitivity

Journal of Biomolecular NMR
Péter KirályGareth A Morris

Abstract

Spectral resolution in proton NMR spectroscopy is reduced by the splitting of resonances into multiplets due to the effect of homonuclear scalar couplings. Although these effects are often hidden in protein NMR spectroscopy by low digital resolution and routine apodization, behind the scenes homonuclear scalar couplings increase spectral overcrowding. The possibilities for biomolecular NMR offered by new pure shift NMR methods are illustrated here. Both resolution and sensitivity are improved, without any increase in experiment time. In these experiments, free induction decays are collected in short bursts of data acquisition, with durations short on the timescale of J-evolution, interspersed with suitable refocusing elements. The net effect is real-time (t 2) broadband homodecoupling, suppressing the multiplet structure caused by proton-proton interactions. The key feature of the refocusing elements is that they discriminate between the resonances of active (observed) and passive (coupling partner) spins. This can be achieved either by using band-selective refocusing or by the BIRD element, in both cases accompanied by a nonselective 180° proton pulse. The latter method selects the active spins based on their one-bond heteronu...Continue Reading

References

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Citations

Mar 5, 2016·Magnetic Resonance in Chemistry : MRC·Veera Mohana Rao KakitaJagadeesh Bharatam
Sep 8, 2015·Chemical Communications : Chem Comm·M ForoozandehG A Morris
Apr 29, 2016·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Veera M R Kakita, Ramakrishna V Hosur
Sep 7, 2016·Journal of Magnetic Resonance·Lukas KaltschneeChristina M Thiele
Oct 12, 2016·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Veera Mohana Rao Kakita, Ramakrishna V Hosur
Mar 14, 2018·Magnetic Resonance in Chemistry : MRC·Josep SauríGary E Martin
Mar 14, 2018·Chemistry : a European Journal·Mohammadali ForoozandehMathias Nilsson
Dec 24, 2017·Magnetic Resonance in Chemistry : MRC·Peter KiralyGareth A Morris
Dec 15, 2017·Magnetic Resonance in Chemistry : MRC·István Timári, Katalin E Kövér
Jun 30, 2019·Metabolites·Abdul-Hamid EmwasDavid S Wishart
Jan 1, 2018·ChemPlusChem·Tamás GyöngyösiKatalin E Kövér
Jan 5, 2019·Analytical Chemistry·István TimáriRafael Brüschweiler

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Methods Mentioned

BETA
NMR

Software Mentioned

BIRD
gHSQC
VnmrJ
BASHD
DOSY
gBASHD
TopSpin
CAWURST

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