Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins

Dominique P. Frueh; Alison Leed; Haribabu Arthanari; Alexander Koglin; Christopher T. Walsh; Gerhard Wagner

doi:10.1007/s10858-009-9372-5

Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in ¹⁵N-¹³C-ILV methyl labeled proteins

Dominique P. Frueh, Alison Leed, Haribabu Arthanari, Alexander Koglin, Christopher T. Walsh, Gerhard Wagner

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

11 Scopus citations

Abstract

We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record ¹³C- and ¹⁵N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that provide cross-peaks in the indirect dimension with low resolution due to limits in experimental time. The technique is particularly advantageous at high field where even longer experimental times would be required for comparable resolution in NOESY-HSQC experiments. The method is demonstrated at 900 MHz and at 750 MHz on 37 and 31 kDa proteins, respectively. The resolution and time saving provided in this experiment was crucial for solving the structures of these two proteins.

Original language	English (US)
Pages (from-to)	311-318
Number of pages	8
Journal	Journal of Biomolecular NMR
Volume	45
Issue number	3
DOIs	https://doi.org/10.1007/s10858-009-9372-5
State	Published - 2009
Externally published	Yes

Keywords

Distance constraints
High-resolution
NOESY
Non-ribosomal peptide synthetases
Nuclear magnetic resonance
Protein structure
Time-shared

ASJC Scopus subject areas

Biochemistry
Spectroscopy

Access to Document

10.1007/s10858-009-9372-5

Cite this

@article{c2ef826be9144af0bd2fe142082b7cb4,

title = "Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins",

abstract = "We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record 13C- and 15N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that provide cross-peaks in the indirect dimension with low resolution due to limits in experimental time. The technique is particularly advantageous at high field where even longer experimental times would be required for comparable resolution in NOESY-HSQC experiments. The method is demonstrated at 900 MHz and at 750 MHz on 37 and 31 kDa proteins, respectively. The resolution and time saving provided in this experiment was crucial for solving the structures of these two proteins.",

keywords = "Distance constraints, High-resolution, NOESY, Non-ribosomal peptide synthetases, Nuclear magnetic resonance, Protein structure, Time-shared",

author = "Frueh, {Dominique P.} and Alison Leed and Haribabu Arthanari and Alexander Koglin and Walsh, {Christopher T.} and Gerhard Wagner",

note = "Funding Information: Acknowledgments This research was supported by the National Institutes of Health (grants GM 47467, CA127990 and RR 00995) and a Human Frontier Science Fellowship to A.K. A.K is currently a Director{\textquoteright}s fellow at the Los Alamos National Laboratory. We thank Dr. Ruma Banerjee for providing the construct for γGCS. We thank two anonymous reviewers for their comments on water suppression.",

year = "2009",

doi = "10.1007/s10858-009-9372-5",

language = "English (US)",

volume = "45",

pages = "311--318",

journal = "Journal of Biomolecular NMR",

issn = "0925-2738",

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T1 - Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins

AU - Frueh, Dominique P.

AU - Leed, Alison

AU - Arthanari, Haribabu

AU - Koglin, Alexander

AU - Walsh, Christopher T.

AU - Wagner, Gerhard

N1 - Funding Information: Acknowledgments This research was supported by the National Institutes of Health (grants GM 47467, CA127990 and RR 00995) and a Human Frontier Science Fellowship to A.K. A.K is currently a Director’s fellow at the Los Alamos National Laboratory. We thank Dr. Ruma Banerjee for providing the construct for γGCS. We thank two anonymous reviewers for their comments on water suppression.

PY - 2009

Y1 - 2009

N2 - We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record 13C- and 15N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that provide cross-peaks in the indirect dimension with low resolution due to limits in experimental time. The technique is particularly advantageous at high field where even longer experimental times would be required for comparable resolution in NOESY-HSQC experiments. The method is demonstrated at 900 MHz and at 750 MHz on 37 and 31 kDa proteins, respectively. The resolution and time saving provided in this experiment was crucial for solving the structures of these two proteins.

AB - We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record 13C- and 15N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that provide cross-peaks in the indirect dimension with low resolution due to limits in experimental time. The technique is particularly advantageous at high field where even longer experimental times would be required for comparable resolution in NOESY-HSQC experiments. The method is demonstrated at 900 MHz and at 750 MHz on 37 and 31 kDa proteins, respectively. The resolution and time saving provided in this experiment was crucial for solving the structures of these two proteins.

KW - Distance constraints

KW - High-resolution

KW - NOESY

KW - Non-ribosomal peptide synthetases

KW - Nuclear magnetic resonance

KW - Protein structure

KW - Time-shared

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