Frequency Selective Heteronuclear Dipolar Recoupling in Rotating Solids:  Accurate13C−15N Distance Measurements in Uniformly13C,15N-labeled Peptides

Jaroniec, Christopher P.; Tounge, Brett A.; Herzfeld, Judith; Griffin, Robert G.

doi:10.1021/ja003266e

Cited by 248 publications

(361 citation statements)

References 75 publications

(130 reference statements)

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“…which may be explained by the librational motion of the 17 O η -1 H η bond vector. The symmetry-based heteronuclear longitudinal two-spin-order recoupling sequences may be readily used in experiments to measure certain internuclear 17 O-1 H distances selectively following the approach established for the widely used REDOR sequence, 87,88 and progress in this direction combined with further improvements in the symmetric sequence design is under way in our laboratory. In addition, the sequences can be incorporated into existing experiments which use rotorencoding of spin-pair coherences in a second time dimension to determine internuclear distances.…”

Section: Resultsmentioning

confidence: 99%

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological ¹⁷O Solid-State NMR

Brinkmann

Kentgens

2006

J. Phys. Chem. B

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In this contribution we present a comprehensive approach to study hydrogen bonding in biological and biomimetic systems through 17 O and 17 O-1 H solid-state NMR combined with density functional theory calculations of 17 O and 1 H NMR parameters. We explore the signal enhancement of 17 O in L-tyrosine‚HCl using repetitive double-frequency swept radio frequency pulses in solid-state NMR. The technique is compatible with high magnetic fields and fast magic-angle spinning of the sample. A maximum enhancement by a factor of 4.3 is obtained in the signal-to-noise ratio of the selectively excited 17 O central transition in a powdered sample of 17 O η -L-tyrosine‚HCl at an external field of 14.1 T and a spinning frequency of 25 kHz. As little as 128 transients lead to meaningful 17 O spectra of the same sample at an external field of 18.8 T and a spinning frequency of 50 kHz. Furthermore we employed supercycled symmetry-based pulse sequences on the protons to achieve heteronuclear longitudinal two-spin-order (I z S z ) recoupling to determine 17 O-1 H distances. These sequences recouple the heteronuclear dipolar 17 O-1 H couplings, where dipolar truncation is absent, while decoupling the homonuclear proton dipolar interactions. They can be applied at fast magic-angle-spinning frequencies up and beyond 50 kHz and are very robust with respect to 17 O quadrupolar couplings and both 17O and 1 H chemical shift anisotropies, which makes them suitable for the use at high external magnetic fields. The method is demonstrated by determining the 17 O η -1 H distance in L-tyrosine‚HCl at a spinning frequency of 50 kHz and an external field of 18.8 T.

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Section: Resultsmentioning

confidence: 99%

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological ¹⁷O Solid-State NMR

Brinkmann

Kentgens

2006

J. Phys. Chem. B

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“…In 15 N-15 N distance measurements, designed to probe the peptide backbone conformation at sites where uniformly labeled residues occurred sequentially, spin polarization was transferred from backbone 15 N nuclei to directly bonded 13 C α nuclei for signal detection using a transferred echo double resonance (TEDOR) sequence (64,65) after the 15 N fpRFDR-CT period, as previously described (66). In 13 C-13 C distance measurements on AMY04, designed to probe the organization of β-sheets in the fibrils, pulsed spin-locked detection was used to improve sensitivity (67).…”

Section: Solid State Nmrmentioning

confidence: 99%

Peptide Conformation and Supramolecular Organization in Amylin Fibrils: Constraints from Solid-State NMR

et al. 2007

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The 37-residue amylin peptide, also known as islet amyloid polypeptide, forms fibrils that are the main peptide or protein component of amyloid that develops in the pancreas of type 2 diabetes patients. Amylin also readily forms amyloid fibrils in vitro that are highly polymorphic under typical experimental conditions. We describe a protocol for the preparation of synthetic amylin fibrils that exhibit a single predominant morphology, which we call a striated ribbon, in electron microscope and atomic force microscope images. Solid state nuclear magnetic resonance (NMR) measurements on a series of isotopically labeled samples indicate a single molecular structure within the striated ribbons. We use scanning transmission electron microscopy and several types of one-dimensional and two-dimensional solid state NMR techniques to obtain constraints on the peptide conformation and supramolecular structure in these amylin fibrils, and derive molecular structural models that are consistent with the experimental data. The basic structural unit in amylin striated ribbons, which we call the protofilament, contains four-layers of parallel β-sheets, formed by two symmetric layers of amylin molecules. The molecular structure of amylin protofilaments in striated ribbons closely resembles the protofilament in amyloid fibrils with similar morphology formed by the 40-residue β-amyloid peptide that is associated with Alzheimer's disease. Keywordsislet amyloid polypeptide; type 2 diabetes; amyloid fibril structure; electron microscopy; atomic force microscopy Amylin, also called islet amyloid polypeptide or IAPP, is a 37-residue peptide that is cosecreted with insulin by the β-cells of pancreas. The normal biological effects of amylin secretion include inhibition of glucagon secretion and reduction of the rate of gastric emptying, effects that contribute to the maintenance of postprandial glucose homeostasis (1). Amylin is the major peptide or protein component of the islet amyloid found in the pancreas of approximately 90% of type 2 (or non-insulin-dependent) diabetes patients (2,3). Fibrillar amylin has been shown * Corresponding author: Dr. Robert Tycko, National Institutes of Health, Building 5, Room 112, Bethesda, MD 20892-0520. phone 301-402-8272. fax 301-496-0825. e-mail robertty@mail.nih.gov.Supporting Information Available HPLC chromatograms from the purification of both reduced and oxidized amylin by reverse-phase chromatography ( Figure S1); FPLC chromatograms from the purification of both human and rodent amylin by size-exclusion chromatography ( Figure S2); 1D 13 C spectra of amylin fibrils with various isotopic labeling patterns, in lyophilized and rehydrated conditions ( Figures S3-S5); Table of 13 C NMR chemical shifts in amylin fibrils, TALOS predictions of backbone torsion angles based on these shifts, and torsion angles determined from 15 N fpRFDR-CT measurements (Table S1). This material is freely available on the World Wide Web at http://www.acs.org. to be toxic to cultured β-cells (4), and has been propo...

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“…In this case, the approach allowed us to facilely observe the labeled backbone 13 C, measure linewidths, and assess the potential for doing a full assignment. Although the linewidths are a few times larger than those observed in U-13 C, 15 N crystalline peptides and proteins, the recent successes in assignment of these crystalline systems suggest that a fairly long sequence in our peptide can be assigned, followed by a full structure determination [5,39,51,[57][58][59].…”

Section: Applications To Membrane-bound Fusion Peptidesmentioning

confidence: 99%

“…Rotational-echo double-resonance spectroscopy (REDOR) is one of the most widely used magic angle spinning (MAS) NMR techniques for analysis of molecular structure in the solid state [1][2][3][4][5][6]. REDOR has found additional application in detection of formation of chemical bonds [7,8] and in resonance assignment of small peptides [9].…”

Section: Introductionmentioning

confidence: 99%

Application of REDOR subtraction for filtered MAS observation of labeled backbone carbons of membrane-bound fusion peptides

Yang

Parkanzky

Bodner

et al. 2002

Journal of Magnetic Resonance

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Clean MAS observation of 13 C-labeled carbons in membrane-bound HIV-1 and influenza fusion peptides was made by using a rotational-echo double-resonance spectroscopy (REDOR) filter of directly bonded 13 C-15 N pairs. The clean filtering achieved with the REDOR approach is superior to filtering done with sample difference spectroscopy. In one labeling approach, the peptide had labels at a single 13 C carbonyl and its directly bonded 15 N. The resulting chemical shift distribution of the filtered signal is used to assess the distribution of local secondary structures at the labeled carbonyl. For the influenza peptide, the Leu-2 carbonyl chemical shift distribution is shown to vary markedly with lipid and detergent composition, as well as peptide:lipid ratio, suggesting that the local peptide structure also has a strong dependence on these factors. Because most carboxylic-and amino-labeled amino acids are commercially available, this REDOR approach should have broad applicability to chemically synthesized peptides as well as bacterially synthesized proteins. In a second labeling approach, the HIV-1 fusion peptide had U-13 C, 15 N labeling over three sequential residues. When a 1.6 ms REDOR dephasing time is used, only backbone 13 C signals are observed. The resulting spectra are used to determine spectral linewidths and to assess feasibility of assignment of uniformly labeled peptide.

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Frequency Selective Heteronuclear Dipolar Recoupling in Rotating Solids: Accurate¹³C−¹⁵N Distance Measurements in Uniformly¹³C,¹⁵N-labeled Peptides

Cited by 248 publications

References 75 publications

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological ¹⁷O Solid-State NMR

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological ¹⁷O Solid-State NMR

Peptide Conformation and Supramolecular Organization in Amylin Fibrils: Constraints from Solid-State NMR

Application of REDOR subtraction for filtered MAS observation of labeled backbone carbons of membrane-bound fusion peptides

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Frequency Selective Heteronuclear Dipolar Recoupling in Rotating Solids: Accurate13C−15N Distance Measurements in Uniformly13C,15N-labeled Peptides

Cited by 248 publications

References 75 publications

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological 17O Solid-State NMR

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological 17O Solid-State NMR

Peptide Conformation and Supramolecular Organization in Amylin Fibrils: Constraints from Solid-State NMR

Application of REDOR subtraction for filtered MAS observation of labeled backbone carbons of membrane-bound fusion peptides

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Frequency Selective Heteronuclear Dipolar Recoupling in Rotating Solids: Accurate¹³C−¹⁵N Distance Measurements in Uniformly¹³C,¹⁵N-labeled Peptides

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological ¹⁷O Solid-State NMR

Sensitivity Enhancement and Heteronuclear Distance Measurements in Biological ¹⁷O Solid-State NMR