Rapid proton-detected NMR assignment for proteins with fast magic angle spinning

Emeline Barbet-Massin; Andrew J. Pell; Joren S. Retel; Loren B. Andreas; Kristaps Jaudzems; W. Trent Franks; Andrew J. Nieuwkoop; Matthias Hiller; Victoria Higman; Paul Guerry; Andrea Bertarello; Michael J. Knight; Michele Felletti; Tanguy Le Marchand; Svetlana Kotelovica; Inara Akopjana; Kaspars Tars; Monica Stoppini; Vittorio Bellotti; Martino Bolognesi; Stefano Ricagno; James J. Chou; Robert G. Griffin; Hartmut Oschkinat; Anne Lesage; Lyndon Emsley; Torsten Herrmann; Guido Pintacuda

doi:10.1021/ja507382j

Rapid proton-detected NMR assignment for proteins with fast magic angle spinning

Emeline Barbet-Massin, Andrew J. Pell, Joren S. Retel, Loren B. Andreas, Kristaps Jaudzems, W. Trent Franks, Andrew J. Nieuwkoop, Matthias Hiller, Victoria Higman, Paul Guerry, Andrea Bertarello, Michael J. Knight, Michele Felletti, Tanguy Le Marchand, Svetlana Kotelovica, Inara Akopjana, Kaspars Tars, Monica Stoppini, Vittorio Bellotti, Martino BolognesiStefano Ricagno, James J. Chou, Robert G. Griffin, Hartmut Oschkinat, Anne Lesage, Lyndon Emsley, Torsten Herrmann, Guido Pintacuda^*

^*Corresponding author for this work

School of Chemistry

Research output: Contribution to journal › Article (Academic Journal) › peer-review

256 Citations (Scopus)

Abstract

Using a set of six ¹H-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide ²H/ ¹H exchange, high magnetic fields, and high-spinning frequencies (ω_r/2π ≥ 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary ¹³C/¹⁵N-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.

Original language	English
Pages (from-to)	12489-12497
Number of pages	9
Journal	Journal of the American Chemical Society
Volume	136
Issue number	35
DOIs	https://doi.org/10.1021/ja507382j
Publication status	Published - 3 Sept 2014

Access to Document

10.1021/ja507382j

Handle.net

Persistent link

Cite this

Barbet-Massin, E., Pell, A. J., Retel, J. S., Andreas, L. B., Jaudzems, K., Franks, W. T., Nieuwkoop, A. J., Hiller, M., Higman, V., Guerry, P., Bertarello, A., Knight, M. J., Felletti, M., Le Marchand, T., Kotelovica, S., Akopjana, I., Tars, K., Stoppini, M., Bellotti, V., ... Pintacuda, G. (2014). Rapid proton-detected NMR assignment for proteins with fast magic angle spinning. Journal of the American Chemical Society, 136(35), 12489-12497. https://doi.org/10.1021/ja507382j

@article{228214dfd7ff413eaed3725fab5b5db4,

title = "Rapid proton-detected NMR assignment for proteins with fast magic angle spinning",

abstract = "Using a set of six 1H-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide 2H/ 1H exchange, high magnetic fields, and high-spinning frequencies (ωr/2π ≥ 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary 13C/15N-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.",

author = "Emeline Barbet-Massin and Pell, {Andrew J.} and Retel, {Joren S.} and Andreas, {Loren B.} and Kristaps Jaudzems and Franks, {W. Trent} and Nieuwkoop, {Andrew J.} and Matthias Hiller and Victoria Higman and Paul Guerry and Andrea Bertarello and Knight, {Michael J.} and Michele Felletti and {Le Marchand}, Tanguy and Svetlana Kotelovica and Inara Akopjana and Kaspars Tars and Monica Stoppini and Vittorio Bellotti and Martino Bolognesi and Stefano Ricagno and Chou, {James J.} and Griffin, {Robert G.} and Hartmut Oschkinat and Anne Lesage and Lyndon Emsley and Torsten Herrmann and Guido Pintacuda",

year = "2014",

month = sep,

day = "3",

doi = "10.1021/ja507382j",

language = "English",

volume = "136",

pages = "12489--12497",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "American Chemical Society",

number = "35",

}

Barbet-Massin, E, Pell, AJ, Retel, JS, Andreas, LB, Jaudzems, K, Franks, WT, Nieuwkoop, AJ, Hiller, M, Higman, V, Guerry, P, Bertarello, A, Knight, MJ, Felletti, M, Le Marchand, T, Kotelovica, S, Akopjana, I, Tars, K, Stoppini, M, Bellotti, V, Bolognesi, M, Ricagno, S, Chou, JJ, Griffin, RG, Oschkinat, H, Lesage, A, Emsley, L, Herrmann, T & Pintacuda, G 2014, 'Rapid proton-detected NMR assignment for proteins with fast magic angle spinning', Journal of the American Chemical Society, vol. 136, no. 35, pp. 12489-12497. https://doi.org/10.1021/ja507382j

TY - JOUR

T1 - Rapid proton-detected NMR assignment for proteins with fast magic angle spinning

AU - Barbet-Massin, Emeline

AU - Pell, Andrew J.

AU - Retel, Joren S.

AU - Andreas, Loren B.

AU - Jaudzems, Kristaps

AU - Franks, W. Trent

AU - Nieuwkoop, Andrew J.

AU - Hiller, Matthias

AU - Higman, Victoria

AU - Guerry, Paul

AU - Bertarello, Andrea

AU - Knight, Michael J.

AU - Felletti, Michele

AU - Le Marchand, Tanguy

AU - Kotelovica, Svetlana

AU - Akopjana, Inara

AU - Tars, Kaspars

AU - Stoppini, Monica

AU - Bellotti, Vittorio

AU - Bolognesi, Martino

AU - Ricagno, Stefano

AU - Chou, James J.

AU - Griffin, Robert G.

AU - Oschkinat, Hartmut

AU - Lesage, Anne

AU - Emsley, Lyndon

AU - Herrmann, Torsten

AU - Pintacuda, Guido

PY - 2014/9/3

Y1 - 2014/9/3

N2 - Using a set of six 1H-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide 2H/ 1H exchange, high magnetic fields, and high-spinning frequencies (ωr/2π ≥ 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary 13C/15N-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.

AB - Using a set of six 1H-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide 2H/ 1H exchange, high magnetic fields, and high-spinning frequencies (ωr/2π ≥ 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary 13C/15N-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.

UR - http://www.scopus.com/inward/record.url?scp=84906862072&partnerID=8YFLogxK

U2 - 10.1021/ja507382j

DO - 10.1021/ja507382j

M3 - Article (Academic Journal)

C2 - 25102442

AN - SCOPUS:84906862072

SN - 0002-7863

VL - 136

SP - 12489

EP - 12497

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 35

ER -

Rapid proton-detected NMR assignment for proteins with fast magic angle spinning

Abstract

Access to Document

Handle.net

Other files and links

Fingerprint

Cite this