Multiple frequency saturation pulses reduce CEST acquisition time for quantifying conformational exchange in biomolecules

Maureen Leninger, William M. Marsiglia, Alexej Jerschow, Nathaniel Traaseth

Research output: Contribution to journalArticle

Abstract

Exchange between conformational states is required for biomolecular catalysis, allostery, and folding. A variety of NMR experiments have been developed to quantify motional regimes ranging from nanoseconds to seconds. In this work, we describe an approach to speed up the acquisition of chemical exchange saturation transfer (CEST) experiments that are commonly used to probe millisecond to second conformational exchange in proteins and nucleic acids. The standard approach is to obtain CEST datasets through the acquisition of a series of 2D correlation spectra where each experiment utilizes a single saturation frequency to 1H, 15N or 13C. These pseudo 3D datasets are time consuming to collect and are further lengthened by reduced signal to noise stemming from the long saturation pulse. In this article, we show how usage of a multiple frequency saturation pulse (i.e., MF-CEST) changes the nature of data collection from series to parallel, and thus decreases the total acquisition time by an integer factor corresponding to the number of frequencies in the pulse. We demonstrate the applicability of MF-CEST on a Src homology 2 (SH2) domain from phospholipase Cγ and the secondary active transport protein EmrE as model systems by collecting 13C methyl and 15N backbone datasets. MF-CEST can also be extended to additional sites within proteins and nucleic acids. The only notable drawback of MF-CEST as applied to backbone 15N experiments occurs when a large chemical shift difference between the major and minor populations is present (typically greater than ~ 8 ppm). In these cases, ambiguity may arise between the chemical shift of the minor population and the multiple frequency saturation pulse. Nevertheless, this drawback does not occur for methyl group MF-CEST experiments or in cases where somewhat smaller chemical shift differences occur are present.

Original languageEnglish (US)
Pages (from-to)19-30
Number of pages12
JournalJournal of Biomolecular NMR
Volume71
Issue number1
DOIs
StatePublished - May 1 2018

Fingerprint

Biomolecules
Nucleic Acids
Chemical shift
src Homology Domains
Active Biological Transport
Type C Phospholipases
Catalysis
Population
Noise
Carrier Proteins
Proteins
Experiments
Datasets
Ion exchange
Nuclear magnetic resonance

Keywords

  • CEST
  • Fast data acquisition
  • Proteins
  • Sensitivity enhancement
  • Solution NMR

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

Cite this

Multiple frequency saturation pulses reduce CEST acquisition time for quantifying conformational exchange in biomolecules. / Leninger, Maureen; Marsiglia, William M.; Jerschow, Alexej; Traaseth, Nathaniel.

In: Journal of Biomolecular NMR, Vol. 71, No. 1, 01.05.2018, p. 19-30.

Research output: Contribution to journalArticle

@article{3a74eb496a784cf2b219d340c7b16999,
title = "Multiple frequency saturation pulses reduce CEST acquisition time for quantifying conformational exchange in biomolecules",
abstract = "Exchange between conformational states is required for biomolecular catalysis, allostery, and folding. A variety of NMR experiments have been developed to quantify motional regimes ranging from nanoseconds to seconds. In this work, we describe an approach to speed up the acquisition of chemical exchange saturation transfer (CEST) experiments that are commonly used to probe millisecond to second conformational exchange in proteins and nucleic acids. The standard approach is to obtain CEST datasets through the acquisition of a series of 2D correlation spectra where each experiment utilizes a single saturation frequency to 1H, 15N or 13C. These pseudo 3D datasets are time consuming to collect and are further lengthened by reduced signal to noise stemming from the long saturation pulse. In this article, we show how usage of a multiple frequency saturation pulse (i.e., MF-CEST) changes the nature of data collection from series to parallel, and thus decreases the total acquisition time by an integer factor corresponding to the number of frequencies in the pulse. We demonstrate the applicability of MF-CEST on a Src homology 2 (SH2) domain from phospholipase Cγ and the secondary active transport protein EmrE as model systems by collecting 13C methyl and 15N backbone datasets. MF-CEST can also be extended to additional sites within proteins and nucleic acids. The only notable drawback of MF-CEST as applied to backbone 15N experiments occurs when a large chemical shift difference between the major and minor populations is present (typically greater than ~ 8 ppm). In these cases, ambiguity may arise between the chemical shift of the minor population and the multiple frequency saturation pulse. Nevertheless, this drawback does not occur for methyl group MF-CEST experiments or in cases where somewhat smaller chemical shift differences occur are present.",
keywords = "CEST, Fast data acquisition, Proteins, Sensitivity enhancement, Solution NMR",
author = "Maureen Leninger and Marsiglia, {William M.} and Alexej Jerschow and Nathaniel Traaseth",
year = "2018",
month = "5",
day = "1",
doi = "10.1007/s10858-018-0186-1",
language = "English (US)",
volume = "71",
pages = "19--30",
journal = "Journal of Biomolecular NMR",
issn = "0925-2738",
publisher = "Springer Netherlands",
number = "1",

}

TY - JOUR

T1 - Multiple frequency saturation pulses reduce CEST acquisition time for quantifying conformational exchange in biomolecules

AU - Leninger, Maureen

AU - Marsiglia, William M.

AU - Jerschow, Alexej

AU - Traaseth, Nathaniel

PY - 2018/5/1

Y1 - 2018/5/1

N2 - Exchange between conformational states is required for biomolecular catalysis, allostery, and folding. A variety of NMR experiments have been developed to quantify motional regimes ranging from nanoseconds to seconds. In this work, we describe an approach to speed up the acquisition of chemical exchange saturation transfer (CEST) experiments that are commonly used to probe millisecond to second conformational exchange in proteins and nucleic acids. The standard approach is to obtain CEST datasets through the acquisition of a series of 2D correlation spectra where each experiment utilizes a single saturation frequency to 1H, 15N or 13C. These pseudo 3D datasets are time consuming to collect and are further lengthened by reduced signal to noise stemming from the long saturation pulse. In this article, we show how usage of a multiple frequency saturation pulse (i.e., MF-CEST) changes the nature of data collection from series to parallel, and thus decreases the total acquisition time by an integer factor corresponding to the number of frequencies in the pulse. We demonstrate the applicability of MF-CEST on a Src homology 2 (SH2) domain from phospholipase Cγ and the secondary active transport protein EmrE as model systems by collecting 13C methyl and 15N backbone datasets. MF-CEST can also be extended to additional sites within proteins and nucleic acids. The only notable drawback of MF-CEST as applied to backbone 15N experiments occurs when a large chemical shift difference between the major and minor populations is present (typically greater than ~ 8 ppm). In these cases, ambiguity may arise between the chemical shift of the minor population and the multiple frequency saturation pulse. Nevertheless, this drawback does not occur for methyl group MF-CEST experiments or in cases where somewhat smaller chemical shift differences occur are present.

AB - Exchange between conformational states is required for biomolecular catalysis, allostery, and folding. A variety of NMR experiments have been developed to quantify motional regimes ranging from nanoseconds to seconds. In this work, we describe an approach to speed up the acquisition of chemical exchange saturation transfer (CEST) experiments that are commonly used to probe millisecond to second conformational exchange in proteins and nucleic acids. The standard approach is to obtain CEST datasets through the acquisition of a series of 2D correlation spectra where each experiment utilizes a single saturation frequency to 1H, 15N or 13C. These pseudo 3D datasets are time consuming to collect and are further lengthened by reduced signal to noise stemming from the long saturation pulse. In this article, we show how usage of a multiple frequency saturation pulse (i.e., MF-CEST) changes the nature of data collection from series to parallel, and thus decreases the total acquisition time by an integer factor corresponding to the number of frequencies in the pulse. We demonstrate the applicability of MF-CEST on a Src homology 2 (SH2) domain from phospholipase Cγ and the secondary active transport protein EmrE as model systems by collecting 13C methyl and 15N backbone datasets. MF-CEST can also be extended to additional sites within proteins and nucleic acids. The only notable drawback of MF-CEST as applied to backbone 15N experiments occurs when a large chemical shift difference between the major and minor populations is present (typically greater than ~ 8 ppm). In these cases, ambiguity may arise between the chemical shift of the minor population and the multiple frequency saturation pulse. Nevertheless, this drawback does not occur for methyl group MF-CEST experiments or in cases where somewhat smaller chemical shift differences occur are present.

KW - CEST

KW - Fast data acquisition

KW - Proteins

KW - Sensitivity enhancement

KW - Solution NMR

UR - http://www.scopus.com/inward/record.url?scp=85047268098&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85047268098&partnerID=8YFLogxK

U2 - 10.1007/s10858-018-0186-1

DO - 10.1007/s10858-018-0186-1

M3 - Article

VL - 71

SP - 19

EP - 30

JO - Journal of Biomolecular NMR

JF - Journal of Biomolecular NMR

SN - 0925-2738

IS - 1

ER -