caa a22 4500 445325976 CHVBK 20180317142720.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s10858-011-9507-3 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9507-3 Improved accuracy in measuring one-bond and two-bond 15N,13Cα coupling constants in proteins by double-inphase/antiphase (DIPAP) spectroscopy [Elektronische Daten] [Frank Löhr, Sina Reckel, Susanne Stefer, Volker Dötsch, Jürgen Schmidt] An extension to HN(CO-α/β-N,Cα-J)-TROSY (Permi and Annila in J Biomol NMR 16:221-227, 2000) is proposed that permits the simultaneous determination of the four coupling constants 1 J N′(i)Cα(i), 2 J HN(i)Cα(i), 2 J Cα(i−1)N′(i), and 3 J Cα(i−1)HN(i) in 15N,13C-labeled proteins. Contrasting the original scheme, in which two separate subspectra exhibit the 2 J CαN′ coupling as inphase and antiphase splitting (IPAP), we here record four subspectra that exhibit all combinations of inphase and antiphase splittings possible with respect to both 2 J CαN′ and 1 J N′Cα (DIPAP). Complementary sign patterns in the different spectrum constituents overdetermine the coupling constants which can thus be extracted at higher accuracy than is possible with the original experiment. Fully exploiting data redundance, simultaneous 2D lineshape fitting of the E.COSY multiplet tilts in all four subspectra provides all coupling constants at ultimate precision. Cross-correlation and differential-relaxation effects were taken into account in the evaluation procedure. By applying a four-point Fourier transform, the set of spectra is reversibly interconverted between DIPAP and spin-state representations. Methods are exemplified using proteins of various size. Springer Science+Business Media B.V., 2011 Correlation nationallicence Fourier transform nationallicence Heteronuclear coupling constant nationallicence Multiplet simulation nationallicence Protein structure nationallicence Spin-state editing nationallicence Ubiquitin nationallicence Ribonuclease T1 nationallicence Xylanase nationallicence Get2 domain nationallicence Proteorhodopsin nationallicence Löhr Frank Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut Reckel Sina Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut Stefer Susanne Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut Dötsch Volker Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut Schmidt Jürgen School of Biosciences, University of Kent, Giles Lane, CT2 7NJ, Canterbury, Kent, UK aut Journal of Biomolecular NMR Springer Netherlands 50/2(2011-06-01), 167-190 0925-2738 50:2<167 2011 50 10858 https://doi.org/10.1007/s10858-011-9507-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9507-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Löhr Frank Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut NATIONALLICENCE 700 1- Reckel Sina Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut NATIONALLICENCE 700 1- Stefer Susanne Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut NATIONALLICENCE 700 1- Dötsch Volker Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe-University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany aut NATIONALLICENCE 700 1- Schmidt Jürgen School of Biosciences, University of Kent, Giles Lane, CT2 7NJ, Canterbury, Kent, UK aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 50/2(2011-06-01), 167-190 0925-2738 50:2<167 2011 50 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer