Improvement of relayed—NOESY type experiments by implantation of spin‐lock sequences

Kessler, Horst; Gemmecker, Gerd; Haase, Burkhard; Steuernagel, Stefan

doi:10.1002/mrc.1260261018

Cited by 16 publications

(13 citation statements)

References 25 publications

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“…For isotropic mixing, either DIPSI-2 (20) at 325 K or MOCCA-XY16 (21) at 253 K pulse sequences were used with duration times 11-100 and 50 ms, respectively. The two-dimensional TOCSY and NOESY spectra were typically acquired with acquisition times of 0.3 and 0.08 s in the direct and indirect dimensions, respectively, with total experiment times of 12 h at 325 K and 48 h at 253 K. The 13 C chemical shifts of ␣ and ␤ carbons and their temperature dependence were measured from two-dimensional 1 H- 13 C HSQC spectra recorded in sensitivity-enhanced mode (22) with 0.085-s acquisition times in both dimensions and total experiment time of 4 -8 h. Spectral assignments were further confirmed and stereo-specific assignments obtained with the help of two-dimensional 1 H-13 C HMBC experiment (23) recorded with 0.14-and 0.085-s acquisition times in direct and indirect dimensions, respectively, for a total acquisition time of 28 h. The 1 -decoupled (24) TOCSY-NOESY spectra (25) were recorded with 1-ms 180° 1 -decoupling pulse and 0.38-and 0.15-s acquisition times in direct and indirect dimensions, respectively, for a total experiment time of 12 h. The H-D exchange was monitored by recording one-dimensional proton spectra after reconstitution of chHepc from H 2 O into D 2 O solutions. Water suppression was achieved by either use of gradients, presaturation pulses, or excitation sculpting.…”

Section: Purification Of Urinary Human Hepcidin (Uhhepc)supporting

confidence: 53%

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Hepcidin Revisited, Disulfide Connectivity, Dynamics, and Structure

Jordan¹,

Poppe²,

Haniu³

et al. 2009

Journal of Biological Chemistry

192

190

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Hepcidin is a tightly folded 25-residue peptide hormone containing four disulfide bonds, which has been shown to act as the principal regulator of iron homeostasis in vertebrates. We used multiple techniques to demonstrate a disulfide bonding pattern for hepcidin different from that previously published. . NMR studies reveal a new model for hepcidin that, at ambient temperatures, interconverts between two different conformations, which could be individually resolved by temperature variation. Using these methods, the solution structure of hepcidin was determined at 325 and 253 K in supercooled water. X-ray analysis of a co-crystal with Fab appeared to stabilize a hepcidin conformation similar to the high temperature NMR structure.

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Section: Purification Of Urinary Human Hepcidin (Uhhepc)supporting

confidence: 53%

“…In contrast, the ␤-proton resonances were highly congested, complicating the observation of H␤-H␤ NOEs. However, in the TOCSY-NOESY relay experiment (25), the H␤-H␤ NOE peaks appear away from the diagonal allowing unambiguous resonance assignment. We used this method to identify Cys 2 H␤-Cys ature limit.…”

mentioning

confidence: 99%

Hepcidin Revisited, Disulfide Connectivity, Dynamics, and Structure

Jordan¹,

Poppe²,

Haniu³

et al. 2009

Journal of Biological Chemistry

192

190

View full text Add to dashboard Cite

show abstract

“…1D TOCSY/HOHAHA [7, 81 spectra were obtained as suggested by Subramanian and Bax [8], with variable mixing time during accumulation; the DANTE pulse sequence [21] was used for selective excitation. The TOR0 experiment [14] was carried out as previously described [I 3, 221. For ROESY spectra [9, 101, the pulse sequence proposed by Rance [23] was applied.…”

Section: Nmr Spectroscopymentioning

confidence: 99%

“…Further propagation of magnetization was halted, however, because of the small 3J4,5 coupling constant. To circumvent this difficulty, we applied a 1D version [13, 211 of the TORO technique [14]. In the first step of the experiment, magnetization was transferred by a through-bond TOCSY mechanism from the irradiated methyl group to H5.…”

Section: N M R Spectramentioning

confidence: 99%

O‐specific polysaccharide of Hafnia alvei lipopolysaccharide isolated from strain 1211

Katzenellenbogen

Romanowska

Dąbrowski

et al. 1991

European Journal of Biochemistry

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The Hajnia alvei strain 121 1 0-specific polysaccharide is composed of 3-amino-N-(~-3'-hydroxybutyryl)-3,6-dideoxy-D-galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine and D-glucose (1 : 1 : 2 : 2). On the basis of sugar and methylation analyses, Smith degradation, and one-and two-dimensional 'H-and I3C-NMR spectroscopy, the polysaccharide was shown to be an 0-acetylated polymer of the repeating hexasaccharide unit, + 2~(4-OAc)Fucp3NAcylj?I + 6~GlcpNAccll --+ (~Glcppl 4 3)4~GalpNAccll 4 3~GlcpNAcBl + 2~Glcppl +, where nFucp3NAcyl= 3-amino-N-(~-3'-hydroxybutyryl)-3,6-dideoxy-~-galactopyranose. The 0-specific polysaccharide showed some microheterogeneity due to incomplete substitution by terminal glucose.Hafnia alvei, a member of Enterobacteriaceae, is an opportunistic pathogen found in some incidences of nosocomial infections [l]. Preliminary chemical characterization of lipopolysaccharides isolated from the H . alvei group has been reported [2]. Recently, we described the structure of 0-specific polysaccharides of four representatives of H. alvei: standard strain ATCC 13337 and strains 1187, 39 and 2 [3-51. As a part of a systematic work, we now report the structural studies on the 0-antigen of H . alvei strain 121 1.The assignment of 'H-NMR signals for the particular sugar residues was performed using two-dimensional (2D) correlation spectroscopy (COSY), 2D relayed coherence transfer (RCT) [6] and one-dimensional (1 D) totally correlated (TOCSY) homonuclear Hartmann-Hahn (HOHAHA) [7, 81 spectroscopy. For sequence and linkage analysis, 2D NOE (NOESY) and rotating frame NOE (ROESY) [9, 101 spectra were recorded. Due to the different substitution pattern of each of the constituent sugar units, these units could be unambiguously identified by comparison of the linkage information obtained from NMR spectroscopy with the data provided by chemical methods and methylation analysis. In this way, we could dispense with the de novo identification of sugar residues by NMR, usually applied in our laboratory [3, 11 -131, which would be difficult in this case in view of seriousCorrespondence to E. Romanowska,

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“…Maximum NOE intensities of several intra-and interglycosidic linkages were obtained with mixing times of 400-500 ms. ROESY-TOCSY (ROTO) 19,20 (TOCSY mixing, time D 150 ms, ROESY mixing time D 300 ms) (Fig. 3) experiments were performed to obtain the NOEs which provided information about the intra-and interglycosidic linkages.…”

Section: Analysis Of Solution Conformation Of Hpg-b2-n5amentioning

confidence: 99%

Determination of solution conformation of allergenically active pentasaccharitol obtained from sea squirt antigen

Kato

Ohta

Munakata

et al. 2001

Magnetic Reson in Chemistry

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Pentasaccharitol, GalNAcb1-4(GalNAca1-2Fuca1-3)GlcNAcb1-3GalNAc-ol, termed HPG-b2-N5a, is an allergenically active O-linked oligosaccharitol derived from a sea squirt H-antigen. Concerning the interaction between the allergen and specific IgE, an analysis of the solution conformation of the saccharitol was carried out. 1 H and 13 C NMR assignments for the pentasaccharitol were obtained using 1D-HOHAHA, TOCSY, DEPT135, HSQC and HMBC techniques. The solution conformation was determined using a combination of 2D-NMR ROESY data and distance geometry (DG) calculations in dihedral angle space with the DADAS 90 program. Two conformers, family A (FamA) and family B (FamB), that satisfied almost all the NOE constraints were obtained, and supplemental MD calculations revealed that FamA was in conformational equilibrium whereas FamB was rather flexible. The molecule of HPG-b2-N5a had a layered and not a linear conformation.

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Improvement of relayed—NOESY type experiments by implantation of spin‐lock sequences

Cited by 16 publications

References 25 publications

Hepcidin Revisited, Disulfide Connectivity, Dynamics, and Structure

Hepcidin Revisited, Disulfide Connectivity, Dynamics, and Structure

O‐specific polysaccharide of Hafnia alvei lipopolysaccharide isolated from strain 1211

Determination of solution conformation of allergenically active pentasaccharitol obtained from sea squirt antigen

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