Selective diagonal-free 13C,13C-edited aliphatic–aromatic NOESY experiment with non-uniform sampling

Abstract

A band-selective aromatic–aliphatic C,C-edited four-dimensional NOESY experiment is proposed here. Its key advantage is the absence of auto-correlation signals which makes it very attractive for joint use with non-uniform sampling. It is demonstrated here that the sensitivity of the experiment is not significantly affected by utilization of selective pulses (for either aromatic-13C or aliphatic-13C spins). The method was applied to the sample of E32Q mutant of human S100A1 protein, a homodimer of total molecular mass ~20 kDa. High-resolution 4D spectra were obtained from ~1.5 % of sampling points required conventionally. It is shown that superior resolution facilitates unambiguous assignment of observed aliphatic–aromatic cross-peaks. Additionally, the addition of aliphatic-13C dimension enables to resolve peaks with degenerated aliphatic 1H chemical shifts. All observed cross-peaks were validated against previously determined 3D structure of E32Q mutant of S100A1 protein (PDB 2LHL). The increased reliability of structural constraints obtained from the proposed high-resolution 4D 13C(ali),13C(aro)-edited NOESY can be exploited in the automated protocols of structure determination of proteins.Electronic supplementary materialThe online version of this article (doi:10.1007/s10858-013-9739-5) contains supplementary material, which is available to authorized users.

15N and 13C- SOFAST-HMQC editing enhances 3D-NOESY sensitivity in highly deuterated, selectively [1H,13C]-labeled proteins

“…The NOE crosspeaks are acquired with high 13 C digital resolution and without autocorrelation and folded peaks. (Stanek et al 2013) The pulse sequence details are given in Fig. 3a and results of the Ernst angle optimization are shown in Supp.…”

“…The example of SOFAST aromatic-methyl NOESY pulse sequence (Fig. 3a) is simpler than the counterparts using HSQC schemes (Morshauser and Zuiderweg 1999; Stanek et al 2013; Xia et al 2001) (Fig. 3b) and scales well to larger methyl-aromatic labeled systems.…”

Applications of high dimensionality experiments to biomolecular NMR

“…This approach is reasonably efficient unless significant overlap of H β and C β in aromatic amino acids occurs. In this case, analysis of 4D 13 C ali , 13 C aroedited HMQC-NOESY-HSQC [132] allows disambiguation of the assignment of correlations between H β and C β and between H δ and C δ .…”

“…4D 13 C ali , 13 C aro -edited HMQC-NOESY-HSQC [132], in which the selective manipulation of 13 C spins with shaped pulses leads to diagonal suppression with a minor loss of sensitivity.…”

The solution structure of the MANEC-type domain from hepatocyte growth factor activator inhibitor-1 reveals an unexpected PAN/apple domain-type fold

“…Aliphatic side chain assignment was achieved using 1 H-13 C HSQC, 4D H ab C ab (CO)NH [22,23] and 4D HCCH-TOCSY [21,24,25]. The aromatic side chain resonances were assigned from the analysis of 1 H-13 C HSQC tuned to aromatic carbons, 2D (HB)CB(CGCD)HD, 2D (HB)CB(CGCDCE)HE, 3D HBCB(CGCD)HD, 3D HBCB(CGCDCE)HE [26] acquired at 800 MHz, 3D 13 C-edited NOESY HSQC tuned to aromatic carbons (measured at 600 MHz) [27] and 800 MHz 4D 13 C ali , 13 C aroedited HMQC-NOESY-HSQC [28]. Distance constraints were obtained at 800 MHz from the 15 N-edited NOESY-HSQC [29], 4D 13 C ali , 13 C aro -edited HMQC-NOESY-HSQC, 4D 13 C ali , 13 C aliedited HMQC-NOESY-HMQC [30,31], and 4D 15 N, 13 C-edited HMQC-NOESY-HSQC [21].…”