Proton‐Detected Solid‐State NMR Spectroscopy of Fibrillar and Membrane Proteins

Linser, Rasmus; Dasari, Muralidhar; Hiller, Matthias; Higman, Victoria Ann; Fink, Uwe; Amo, Juan Miguel López del; Marković, Stefan; Handel, Liselotte; Keßler, Brigitte; Schmieder, Peter; Oesterhelt, Dieter; Oschkinat, Hartmut; Reif, Bernd

doi:10.1002/anie.201008244

Cited by 185 publications

(177 citation statements)

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“…This interpretation is consistent with the observation that the N-terminus of the peptide in the amyloid fibril is not structured. 11,23,47 Additional correlations involving the hydroxyl group of Ser-26 and arginine/tyrosine residues are observed, indicating that also these side chains are involved in tertiary contacts.…”

Section: Resultsmentioning

confidence: 99%

“…Amide backbone resonances have been assigned previously. 23 In this manuscript, emphasis is put on the characterization of side chain exchangeable groups, in particular on the histidine resonances.…”

Section: Resultsmentioning

confidence: 99%

“…However, spectra are not easily superimposable even taking the deuterium isotope induced chemical shift changes into account. As in the case for assignment of the backbone amide resonances in perdeuterated Ab fibrils, 23 only one set of resonances is observed. We attribute this observation to a differential thermodynamic stability of protonated and deuterated amyloid fibrils.…”

Section: Discussionmentioning

confidence: 95%

“…14 In all structural models, Ab peptides are arranged in a parallel fashion. An antiparallel arrangement is only observed for the Iowa mutant Ab-D23N, 15 and for truncated forms of the peptide such as Ab [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25] . 16 The fibril structure is stabilized in particular by hydrogen bonding interactions.…”

mentioning

confidence: 99%

“…We and others could show in the past that high resolution proton spectra can be obtained for crystalline proteins, [17][18][19][20][21][22] as well as for amyloid fibrils and membrane proteins. 23,24 Due to the fact that magnetization transfer via dipolar interactions is very efficient, quickly exchanging side chain hydroxyl protons can be observed and assigned easily. 25 At the same time, scalar coupling based experiments yield reliable resonance assignments in the solid-state.…”

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confidence: 99%

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Hydrogen bonding involving side chain exchangeable groups stabilizes amyloid quarternary structure

Agarwal

Linser

Dasari

et al. 2013

Phys. Chem. Chem. Phys.

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The amyloid b-peptide (Ab) is the major structural component of amyloid fibrils in the plaques of brains of Alzheimer's disease patients. Numerous studies have addressed important aspects of secondary and tertiary structure of fibrils. In electron microscopic images, fibrils often bundle together. The mechanisms which drive the association of protofilaments into bundles of fibrils are not known. We show here that amino acid side chain exchangeable groups like e.g. histidines can provide useful restraints to determine the quarternary assembly of an amyloid fibril. Exchangeable protons are only observable if a side chain hydrogen bond is formed and the respective protons are protected from exchange. The method relies on deuteration of the Ab peptide. Exchangeable deuterons are substituted with protons, before fibril formation is initiated.Alzheimer's disease (AD) is a slowly progressive neurological disorder which is associated with memory loss, cognitive impairment and finally dementia and death. The amyloid b-peptide (Ab) is the major structural component of amyloid fibrils in the plaques of brains of Alzheimer's disease patients. Ab is a cleavage product resulting from Alzheimer Precursor Protein (APP) processing. 1 The g-secretase cut yields Ab fragments of different lengths of which Ab40 and Ab42 are most abundant. 2,3 In vitro, the Ab peptide aggregates over time into oligomers and fibrillar structures. 4 In the past, several Ab fibril structure models have been suggested. These are based on MAS solidstate NMR experiments, 5-11 and cryo-electron microscopic image reconstructions. 12 Even though there is some controversy concerning the conformational space that Ab fibrils can adopt, all published models, including the 2-fold 7,10 and 3-fold 9 symmetric Ab 1-40 fibril structure suggested by Tycko, and Bertini and co-workers, agree on the basic building block which involves a b-sheet (b1, residues 12-24), a turn, and a second b-sheet (b2, residues 28-40). Biophysical studies indicate a certain degree of conformational plasticity for the N-terminal b-sheet. 13,14 Whereas b2 is present in all the studies, b1 can be truncated or does not adopt a regular b-sheet structure. Depending on the preparation conditions, amide protons in the region of the peptide where the first b-strand is expected show differential H/D protection factors. 13 Similarly, a certain degree of conformational variability in the N-terminus is suggested using cryo-electron microscopy. 14 In all structural models, Ab peptides are arranged in a parallel fashion. An antiparallel arrangement is only observed for the Iowa mutant Ab-D23N, 15 and for truncated forms of the peptide such as Ab [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25] . 16 The fibril structure is stabilized in particular by hydrogen bonding interactions. Therefore, observation of exchangeable and titratable groups is of particular interest to identify hydrogen bonding patterns. We and others could show in the past that high resolution proton spectra can be obtained for...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 95%