Measurement ofJand Dipolar Couplings from Simplified Two-Dimensional NMR Spectra

Ottiger, Marcel; Delaglio, Frank; Bax, Ad

doi:10.1006/jmre.1998.1361

Cited by 939 publications

(783 citation statements)

References 19 publications

(17 reference statements)

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“…Two-dimensional heteronuclear single quantum coherence experiments were recorded without 1 H decoupling and both in bulk and in the presence of the gel, using the IPAP sequence [12]. 15 N-Edited nuclear Overhauser spectroscopy (NOESY) were run as described in [13].…”

Section: Methodsmentioning

confidence: 99%

NMR studies of flexible peptides in cavities mimicking the synaptic cleft

et al. 2002

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The interaction of neuropeptides with post-synaptic receptors is characterised by a high entropic barrier originating from the combination of nanomolar concentration with low conformer population. The influence of high viscosity environments on conformer distribution can help overcome this difficulty. In an attempt to simulate the physicochemical conditions of the synaptic cleft, 15 N-labelled enkephalin has been studied in polyacrylamide gels swollen by different aqueous solutions in the temperature range 273^293 K. Nuclear Overhauser enhancement spectra in the gel pores are consistent with a conformational selection or a slowing down of internal motions that can favour the interaction of the peptide with the receptor. ß

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Section: Methodsmentioning

confidence: 99%

NMR studies of flexible peptides in cavities mimicking the synaptic cleft

et al. 2002

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“…The 1 H-15 N RDCs were measured using a sensitivity-enhanced 1 H-15 N IPAP (in-phaseanti-phase) experiment modified for suppression of the NH 2 signals from the acryl-amide in the gel [46,48,49]. The contribution to the residual dipolar coupling splitting from the isotropic scalar coupling was determined by performing the same experiment on an isotropic micelle sample, and subtracting the value of the isotropic J-coupling obtained from that measured for the weakly aligned gel sample.…”

Section: Solution Nmr Structural Studies In Micellesmentioning

confidence: 99%

NMR of membrane proteins in micelles and bilayers: The FXYD family proteins

Franzin

Gong

Thai

et al. 2007

Methods

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Determining the atomic resolution structures of membrane proteins is of particular interest in contemporary structural biology. Helical membrane proteins constitute one-third of the expressed proteins encoded in a genome, many drugs have membrane-bound proteins as their receptors, and mutations in membrane proteins result in human diseases. Although integral membrane proteins provide daunting technical challenges for all methods of protein structure determination, nuclear magnetic resonance (NMR) spectroscopy can be an extremely versatile and powerful method for determining their structures and characterizing their dynamics, in lipid environments that closely mimic the cell membranes. Once milligram amounts of isotopically labeled protein are expressed and purified, micelle samples can be prepared for solution NMR analysis, and lipid bilayer samples can be prepared for solid-state NMR analysis. The two approaches are complementary and can provide detailed structural and dynamic information. This paper describes the steps for membrane protein structure determination using solution and solid-state NMR. The methods for protein expression and purification, sample preparation and NMR experiments are described and illustrated with examples from the FXYD proteins, a family of regulatory subunits of the Na, KATPase.

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“…The one bond NÀ ÀH couplings in the isotropic and anisotropic media were measured using two-dimensional IPAP 1 H-15 N HSQC experiments. 19 The amide resonances were successfully assigned, based on those of the NMR sample with 90 mM NaCl. Two sets of { 1 H}-15 N NOE spectra with and without the NOE effect were recorded and analyzed, as described.…”

Section: Nmr Spectroscopymentioning

confidence: 99%