A Suite of Triple Resonance NMR Experiments for the Backbone Assignment of 15N, 13C, 2H Labeled Proteins with High Sensitivity

Toshio Yamazaki, D. R. Muhandiranv, Lewis E. Kay, Weon Tae Lee, Cheryl H. Arrowsmith

Research output: Contribution to journalArticle

456 Citations (Scopus)

Abstract

A suite of NMR pulse schemes is presented for the assignment of HN, 15N, 13Cα, and 13Cβ resonances in 15N, 13C, 2H labeled proteins. The experiments exploit the line narrowing of the 13C spins caused by the substitution of deuterons for bound protons and are therefore well suited for application to molecules with masses on the order of 30-40 kDa where standard triple resonance experiments may fail completely or provide spectra of poor sensitivity. The methods are demonstrated on a 37 kDa ternary complex of a 15N, 13C, and ~70% 2H labeled sample of tip-repressor, the corepressor 5-methyltryptophan, and a 20 basepair trp-operator DNA fragment. All of the experiments in the family presented provide well over 95% of the expected intra-and/or inter-residue correlations. Carbon relaxation measurements of the complex indicate that the use of deuteration increases the average 13Cα T2 value from 16.5 to 130 ms.

Original languageEnglish
Pages (from-to)11655-11666
Number of pages12
JournalJournal of the American Chemical Society
Volume116
Issue number26
DOIs
Publication statusPublished - 1994 Dec 1

Fingerprint

Co-Repressor Proteins
Deuterium
Protons
Carbon
Nuclear magnetic resonance
Proteins
DNA
Experiments
Substitution reactions
Molecules
5-methyltryptophan

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Yamazaki, Toshio ; Muhandiranv, D. R. ; Kay, Lewis E. ; Lee, Weon Tae ; Arrowsmith, Cheryl H. / A Suite of Triple Resonance NMR Experiments for the Backbone Assignment of 15N, 13C, 2H Labeled Proteins with High Sensitivity. In: Journal of the American Chemical Society. 1994 ; Vol. 116, No. 26. pp. 11655-11666.
@article{31de9d5212c541faae76920d5188c599,
title = "A Suite of Triple Resonance NMR Experiments for the Backbone Assignment of 15N, 13C, 2H Labeled Proteins with High Sensitivity",
abstract = "A suite of NMR pulse schemes is presented for the assignment of HN, 15N, 13Cα, and 13Cβ resonances in 15N, 13C, 2H labeled proteins. The experiments exploit the line narrowing of the 13C spins caused by the substitution of deuterons for bound protons and are therefore well suited for application to molecules with masses on the order of 30-40 kDa where standard triple resonance experiments may fail completely or provide spectra of poor sensitivity. The methods are demonstrated on a 37 kDa ternary complex of a 15N, 13C, and ~70{\%} 2H labeled sample of tip-repressor, the corepressor 5-methyltryptophan, and a 20 basepair trp-operator DNA fragment. All of the experiments in the family presented provide well over 95{\%} of the expected intra-and/or inter-residue correlations. Carbon relaxation measurements of the complex indicate that the use of deuteration increases the average 13Cα T2 value from 16.5 to 130 ms.",
author = "Toshio Yamazaki and Muhandiranv, {D. R.} and Kay, {Lewis E.} and Lee, {Weon Tae} and Arrowsmith, {Cheryl H.}",
year = "1994",
month = "12",
day = "1",
doi = "10.1021/ja00105a005",
language = "English",
volume = "116",
pages = "11655--11666",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "26",

}

A Suite of Triple Resonance NMR Experiments for the Backbone Assignment of 15N, 13C, 2H Labeled Proteins with High Sensitivity. / Yamazaki, Toshio; Muhandiranv, D. R.; Kay, Lewis E.; Lee, Weon Tae; Arrowsmith, Cheryl H.

In: Journal of the American Chemical Society, Vol. 116, No. 26, 01.12.1994, p. 11655-11666.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A Suite of Triple Resonance NMR Experiments for the Backbone Assignment of 15N, 13C, 2H Labeled Proteins with High Sensitivity

AU - Yamazaki, Toshio

AU - Muhandiranv, D. R.

AU - Kay, Lewis E.

AU - Lee, Weon Tae

AU - Arrowsmith, Cheryl H.

PY - 1994/12/1

Y1 - 1994/12/1

N2 - A suite of NMR pulse schemes is presented for the assignment of HN, 15N, 13Cα, and 13Cβ resonances in 15N, 13C, 2H labeled proteins. The experiments exploit the line narrowing of the 13C spins caused by the substitution of deuterons for bound protons and are therefore well suited for application to molecules with masses on the order of 30-40 kDa where standard triple resonance experiments may fail completely or provide spectra of poor sensitivity. The methods are demonstrated on a 37 kDa ternary complex of a 15N, 13C, and ~70% 2H labeled sample of tip-repressor, the corepressor 5-methyltryptophan, and a 20 basepair trp-operator DNA fragment. All of the experiments in the family presented provide well over 95% of the expected intra-and/or inter-residue correlations. Carbon relaxation measurements of the complex indicate that the use of deuteration increases the average 13Cα T2 value from 16.5 to 130 ms.

AB - A suite of NMR pulse schemes is presented for the assignment of HN, 15N, 13Cα, and 13Cβ resonances in 15N, 13C, 2H labeled proteins. The experiments exploit the line narrowing of the 13C spins caused by the substitution of deuterons for bound protons and are therefore well suited for application to molecules with masses on the order of 30-40 kDa where standard triple resonance experiments may fail completely or provide spectra of poor sensitivity. The methods are demonstrated on a 37 kDa ternary complex of a 15N, 13C, and ~70% 2H labeled sample of tip-repressor, the corepressor 5-methyltryptophan, and a 20 basepair trp-operator DNA fragment. All of the experiments in the family presented provide well over 95% of the expected intra-and/or inter-residue correlations. Carbon relaxation measurements of the complex indicate that the use of deuteration increases the average 13Cα T2 value from 16.5 to 130 ms.

UR - http://www.scopus.com/inward/record.url?scp=0028578764&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028578764&partnerID=8YFLogxK

U2 - 10.1021/ja00105a005

DO - 10.1021/ja00105a005

M3 - Article

AN - SCOPUS:0028578764

VL - 116

SP - 11655

EP - 11666

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 26

ER -