Optimization of 1H decoupling eliminates sideband artifacts in 3D TROSY-based triple resonance experiments

Youlin Xia; Paolo Rossi; Marco Tonelli; Chengdong Huang; Charalampos G. Kalodimos; Gianluigi Veglia

doi:10.1007/s10858-017-0133-6

Optimization of ¹H decoupling eliminates sideband artifacts in 3D TROSY-based triple resonance experiments

Youlin Xia, Paolo Rossi, Marco Tonelli, Chengdong Huang, Charalampos G. Kalodimos, Gianluigi Veglia

Biochemistry, Molecular Biology, and Biophysics (TMED)

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

TROSY-based triple resonance experiments are essential for protein backbone assignment of large biomolecular systems by solution NMR spectroscopy. In a survey of the current Bruker pulse sequence library for TROSY-based experiments we found that several sequences were plagued by artifacts that affect spectral quality and hamper data analysis. Specifically, these experiments produce sidebands in the ¹³C(t₁) dimension with inverted phase corresponding to ¹H_N resonance frequencies, with approximately 5% intensity of the parent ¹³C crosspeaks. These artifacts originate from the modulation of the ¹H_N frequency onto the resonance frequency of ¹³Cα and/or ¹³Cβ and are due to 180° pulses imperfections used for ¹H decoupling during the ¹³C(t₁) evolution period. These sidebands can become severe for CA_i, CA_i−1 and/or CB_i, CB_i−1 correlation experiments such as TROSY-HNCACB. Here, we implement three alternative decoupling strategies that suppress these artifacts and, depending on the scheme employed, boost the sensitivity up to 14% on Bruker spectrometers. A class of comparable Agilent/Varian pulse sequences that use WALTZ16 ¹H decoupling can also be improved by this method resulting in up to 60–80% increase in sensitivity.

Original language	English (US)
Pages (from-to)	45-52
Number of pages	8
Journal	Journal of biomolecular NMR
Volume	69
Issue number	1
DOIs	https://doi.org/10.1007/s10858-017-0133-6
State	Published - Sep 1 2017

Bibliographical note

Funding Information:
This work is financially supported by the NIH grants GM 100310 to G. V. and AI094623 to C.G.K. The experiments were carried out at the Minnesota NMR Center (MNMR) and at the National Magnetic Resonance Facility at Madison (NMRFAM) [NIH support: P41GM103399 (formerly P41RR002301); P41GM103399, S10RR02781, S10RR08438, S10RR023438, S10RR025062, S10RR029220. NSF support: DMB-8415048, OIA-9977486, BIR-9214394]. Many thanks to Prof. E. Komives and Dr. T. Kromann-Tofting at UCSD for providing testing sample. Youlin Xia and Paolo Rossi have contributed equally to the work.

Funding Information:
Acknowledgements This work is financially supported by the NIH grants GM 100310 to G. V. and AI094623 to C.G.K.. The experiments were carried out at the Minnesota NMR Center (MNMR) and at the National Magnetic Resonance Facility at Madison (NMRFAM) [NIH support: P41GM103399 (formerly P41RR002301); P41GM103399, S10RR02781, S10RR08438, S10RR023438, S10RR025062, S10RR029220. NSF support: DMB-8415048, OIA-9977486, BIR-9214394]. Many thanks to Prof. E. Komives and Dr. T. Kromann-Tofting at UCSD for providing testing sample.

Publisher Copyright:
© 2017, Springer Science+Business Media B.V.

Keywords

Backbone assignment
NMR
Pulse Imperfection
Sideband artifacts
TROSY-based triple resonance

Access

10.1007/s10858-017-0133-6

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5693675

OpenUrl availability

Full text

Cite this

@article{a4625ce24f734fa9aca51be22a56c93f,

title = "Optimization of 1H decoupling eliminates sideband artifacts in 3D TROSY-based triple resonance experiments",

abstract = "TROSY-based triple resonance experiments are essential for protein backbone assignment of large biomolecular systems by solution NMR spectroscopy. In a survey of the current Bruker pulse sequence library for TROSY-based experiments we found that several sequences were plagued by artifacts that affect spectral quality and hamper data analysis. Specifically, these experiments produce sidebands in the 13C(t1) dimension with inverted phase corresponding to 1HN resonance frequencies, with approximately 5% intensity of the parent 13C crosspeaks. These artifacts originate from the modulation of the 1HN frequency onto the resonance frequency of 13Cα and/or 13Cβ and are due to 180° pulses imperfections used for 1H decoupling during the 13C(t1) evolution period. These sidebands can become severe for CAi, CAi−1 and/or CBi, CBi−1 correlation experiments such as TROSY-HNCACB. Here, we implement three alternative decoupling strategies that suppress these artifacts and, depending on the scheme employed, boost the sensitivity up to 14% on Bruker spectrometers. A class of comparable Agilent/Varian pulse sequences that use WALTZ16 1H decoupling can also be improved by this method resulting in up to 60–80% increase in sensitivity.",

keywords = "Backbone assignment, NMR, Pulse Imperfection, Sideband artifacts, TROSY-based triple resonance",

author = "Youlin Xia and Paolo Rossi and Marco Tonelli and Chengdong Huang and Kalodimos, {Charalampos G.} and Gianluigi Veglia",

note = "Funding Information: This work is financially supported by the NIH grants GM 100310 to G. V. and AI094623 to C.G.K. The experiments were carried out at the Minnesota NMR Center (MNMR) and at the National Magnetic Resonance Facility at Madison (NMRFAM) [NIH support: P41GM103399 (formerly P41RR002301); P41GM103399, S10RR02781, S10RR08438, S10RR023438, S10RR025062, S10RR029220. NSF support: DMB-8415048, OIA-9977486, BIR-9214394]. Many thanks to Prof. E. Komives and Dr. T. Kromann-Tofting at UCSD for providing testing sample. Youlin Xia and Paolo Rossi have contributed equally to the work. Funding Information: Acknowledgements This work is financially supported by the NIH grants GM 100310 to G. V. and AI094623 to C.G.K.. The experiments were carried out at the Minnesota NMR Center (MNMR) and at the National Magnetic Resonance Facility at Madison (NMRFAM) [NIH support: P41GM103399 (formerly P41RR002301); P41GM103399, S10RR02781, S10RR08438, S10RR023438, S10RR025062, S10RR029220. NSF support: DMB-8415048, OIA-9977486, BIR-9214394]. Many thanks to Prof. E. Komives and Dr. T. Kromann-Tofting at UCSD for providing testing sample. Publisher Copyright: {\textcopyright} 2017, Springer Science+Business Media B.V.",

year = "2017",

month = sep,

day = "1",

doi = "10.1007/s10858-017-0133-6",

language = "English (US)",

volume = "69",

pages = "45--52",

journal = "Journal of biomolecular NMR",

issn = "0925-2738",

publisher = "Springer Netherlands",

number = "1",

}

TY - JOUR

T1 - Optimization of 1H decoupling eliminates sideband artifacts in 3D TROSY-based triple resonance experiments

AU - Xia, Youlin

AU - Rossi, Paolo

AU - Tonelli, Marco

AU - Huang, Chengdong

AU - Kalodimos, Charalampos G.

AU - Veglia, Gianluigi

N1 - Funding Information: This work is financially supported by the NIH grants GM 100310 to G. V. and AI094623 to C.G.K. The experiments were carried out at the Minnesota NMR Center (MNMR) and at the National Magnetic Resonance Facility at Madison (NMRFAM) [NIH support: P41GM103399 (formerly P41RR002301); P41GM103399, S10RR02781, S10RR08438, S10RR023438, S10RR025062, S10RR029220. NSF support: DMB-8415048, OIA-9977486, BIR-9214394]. Many thanks to Prof. E. Komives and Dr. T. Kromann-Tofting at UCSD for providing testing sample. Youlin Xia and Paolo Rossi have contributed equally to the work. Funding Information: Acknowledgements This work is financially supported by the NIH grants GM 100310 to G. V. and AI094623 to C.G.K.. The experiments were carried out at the Minnesota NMR Center (MNMR) and at the National Magnetic Resonance Facility at Madison (NMRFAM) [NIH support: P41GM103399 (formerly P41RR002301); P41GM103399, S10RR02781, S10RR08438, S10RR023438, S10RR025062, S10RR029220. NSF support: DMB-8415048, OIA-9977486, BIR-9214394]. Many thanks to Prof. E. Komives and Dr. T. Kromann-Tofting at UCSD for providing testing sample. Publisher Copyright: © 2017, Springer Science+Business Media B.V.

PY - 2017/9/1

Y1 - 2017/9/1

N2 - TROSY-based triple resonance experiments are essential for protein backbone assignment of large biomolecular systems by solution NMR spectroscopy. In a survey of the current Bruker pulse sequence library for TROSY-based experiments we found that several sequences were plagued by artifacts that affect spectral quality and hamper data analysis. Specifically, these experiments produce sidebands in the 13C(t1) dimension with inverted phase corresponding to 1HN resonance frequencies, with approximately 5% intensity of the parent 13C crosspeaks. These artifacts originate from the modulation of the 1HN frequency onto the resonance frequency of 13Cα and/or 13Cβ and are due to 180° pulses imperfections used for 1H decoupling during the 13C(t1) evolution period. These sidebands can become severe for CAi, CAi−1 and/or CBi, CBi−1 correlation experiments such as TROSY-HNCACB. Here, we implement three alternative decoupling strategies that suppress these artifacts and, depending on the scheme employed, boost the sensitivity up to 14% on Bruker spectrometers. A class of comparable Agilent/Varian pulse sequences that use WALTZ16 1H decoupling can also be improved by this method resulting in up to 60–80% increase in sensitivity.

AB - TROSY-based triple resonance experiments are essential for protein backbone assignment of large biomolecular systems by solution NMR spectroscopy. In a survey of the current Bruker pulse sequence library for TROSY-based experiments we found that several sequences were plagued by artifacts that affect spectral quality and hamper data analysis. Specifically, these experiments produce sidebands in the 13C(t1) dimension with inverted phase corresponding to 1HN resonance frequencies, with approximately 5% intensity of the parent 13C crosspeaks. These artifacts originate from the modulation of the 1HN frequency onto the resonance frequency of 13Cα and/or 13Cβ and are due to 180° pulses imperfections used for 1H decoupling during the 13C(t1) evolution period. These sidebands can become severe for CAi, CAi−1 and/or CBi, CBi−1 correlation experiments such as TROSY-HNCACB. Here, we implement three alternative decoupling strategies that suppress these artifacts and, depending on the scheme employed, boost the sensitivity up to 14% on Bruker spectrometers. A class of comparable Agilent/Varian pulse sequences that use WALTZ16 1H decoupling can also be improved by this method resulting in up to 60–80% increase in sensitivity.

KW - Backbone assignment

KW - NMR

KW - Pulse Imperfection

KW - Sideband artifacts

KW - TROSY-based triple resonance

UR - http://www.scopus.com/inward/record.url?scp=85028960634&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85028960634&partnerID=8YFLogxK

U2 - 10.1007/s10858-017-0133-6

DO - 10.1007/s10858-017-0133-6

M3 - Article

C2 - 28887770

AN - SCOPUS:85028960634

SN - 0925-2738

VL - 69

SP - 45

EP - 52

JO - Journal of biomolecular NMR

JF - Journal of biomolecular NMR

IS - 1

ER -