Paramagnetic relaxation enhancement to improve sensitivity of fast NMR methods: application to intrinsically disordered proteins.
We report enhanced sensitivity NMR measurements of intrinsically disordered proteins in the presence of paramagnetic relaxation enhancement (PRE) agents such as Ni(2+)-chelated DO2A. In proton-detected (1)H-(15)N SOFAST-HMQC and carbon-detected (H-flip)(13)CO-(15)N experiments, faster longitudinal relaxation enables the usage of even shorter interscan delays. This results in higher NMR signal intensities per units of experimental time, without adverse line broadening effects. At 40 mmol·L(-1) of the PRE agent, we obtain a 1.7- to 1.9-fold larger signal to noise (S/N) for the respective 2D NMR experiments. High solvent accessibility of intrinsically disordered protein (IDP) residues renders this class of proteins particularly amenable to the outlined approach.
Department of NMR-assisted Structural Biology, In-cell NMR Group, Leibniz Institute of Molecular Pharmacology (FMP), Berlin, Germany.
François-Xavier
FX
Theillet
Andres
A
Binolfi
Stamatis
S
Liokatis
Silvia
S
Verzini
Philipp
P
Selenko
D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D009532:Nickel / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity
C561329:1,4,7,10-tetraazacyclododecane-1,7-diacetic acid; D006573:Heterocyclic Compounds, 1-Ring; D011506:Proteins; D009532:Nickel