Saturation-Transfer Difference (STD) NMR: A Simple and Fast Method for Ligand Screening and Characterization of Protein Binding

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

Saturation transfer difference (STD) NMR has emerged as one of the most popular ligand-based NMR techniques for the study of protein ligand interactions. The success of this technique is a consequence of its robustness and the fact that it is focused on the signals of the ligand, without any need of processing NMR information about the receptor and only using small quantities of nonlabeled macromolecule. Moreover, the attractiveness of this experiment is also extendable to the classroom. In the context of a practical NMR class, this experiment is ideal to illustrate some fundamental NMR concepts, such as the nuclear Overhauser effect and relaxation in a multidisciplinary context, bridging chemistry and biochemistry with a taste of medicinal chemistry. We use the readily available human serum albumin (HSA), 6-D,L-methyl-tryptophan (6-CH3-Trp), and 7- D,L-methyl-tryptophan (7-CH3-Trp) to introduce the STD-NMR experiment and to illustrate its applicability for ligand screening, mapping of binding moieties, and determination of the dissociation constant, in a context that can be explored or adapted to the student's course level and topic (chemistry or biochemistry). We also cover the most important theoretical aspects of the STD experiment, calling attention to some of its limitations and drawbacks without a complex theoretical approach.
Original languageEnglish
Pages (from-to)990-994
JournalJournal of Chemical Education
Volume88
Issue number7
DOIs
Publication statusPublished - 18 Apr 2011

Fingerprint

Screening
Nuclear magnetic resonance
Ligands
biochemistry
chemistry
experiment
Biochemistry
Experiments
social attraction
information processing
Macromolecules
Serum Albumin
classroom
Protein Binding
interaction
Students
student
Processing
Proteins
tryptophan methyl ester

Cite this

@article{76c3506d564144538a88e1ab0b92c1f9,
title = "Saturation-Transfer Difference (STD) NMR: A Simple and Fast Method for Ligand Screening and Characterization of Protein Binding",
abstract = "Saturation transfer difference (STD) NMR has emerged as one of the most popular ligand-based NMR techniques for the study of protein ligand interactions. The success of this technique is a consequence of its robustness and the fact that it is focused on the signals of the ligand, without any need of processing NMR information about the receptor and only using small quantities of nonlabeled macromolecule. Moreover, the attractiveness of this experiment is also extendable to the classroom. In the context of a practical NMR class, this experiment is ideal to illustrate some fundamental NMR concepts, such as the nuclear Overhauser effect and relaxation in a multidisciplinary context, bridging chemistry and biochemistry with a taste of medicinal chemistry. We use the readily available human serum albumin (HSA), 6-D,L-methyl-tryptophan (6-CH3-Trp), and 7- D,L-methyl-tryptophan (7-CH3-Trp) to introduce the STD-NMR experiment and to illustrate its applicability for ligand screening, mapping of binding moieties, and determination of the dissociation constant, in a context that can be explored or adapted to the student's course level and topic (chemistry or biochemistry). We also cover the most important theoretical aspects of the STD experiment, calling attention to some of its limitations and drawbacks without a complex theoretical approach.",
keywords = "Spectroscopy, ligands, high-affinity, Upper-Division, Interdisciplinary/Multidisciplinary, Biochemistry, Bioanalytical, Laboratory, Education/Research, Instruction, Learning/Manipulatives, nuclear-magnetic-resonance, Chemistry, NMR, Hands-On, Proteins/Peptides, spectroscopy, Graduate, Drugs/Pharmaceuticals, Undergraduate",
author = "Aldino Viegas and Jo{\~a}o Manso and Cabrita, {Eurico Jos{\'e} da Silva}",
year = "2011",
month = "4",
day = "18",
doi = "10.1021/ed101169t",
language = "English",
volume = "88",
pages = "990--994",
journal = "Journal of Chemical Education",
issn = "0021-9584",
publisher = "AMER CHEMICAL SOC",
number = "7",

}

TY - JOUR

T1 - Saturation-Transfer Difference (STD) NMR: A Simple and Fast Method for Ligand Screening and Characterization of Protein Binding

AU - Viegas, Aldino

AU - Manso, João

AU - Cabrita, Eurico José da Silva

PY - 2011/4/18

Y1 - 2011/4/18

N2 - Saturation transfer difference (STD) NMR has emerged as one of the most popular ligand-based NMR techniques for the study of protein ligand interactions. The success of this technique is a consequence of its robustness and the fact that it is focused on the signals of the ligand, without any need of processing NMR information about the receptor and only using small quantities of nonlabeled macromolecule. Moreover, the attractiveness of this experiment is also extendable to the classroom. In the context of a practical NMR class, this experiment is ideal to illustrate some fundamental NMR concepts, such as the nuclear Overhauser effect and relaxation in a multidisciplinary context, bridging chemistry and biochemistry with a taste of medicinal chemistry. We use the readily available human serum albumin (HSA), 6-D,L-methyl-tryptophan (6-CH3-Trp), and 7- D,L-methyl-tryptophan (7-CH3-Trp) to introduce the STD-NMR experiment and to illustrate its applicability for ligand screening, mapping of binding moieties, and determination of the dissociation constant, in a context that can be explored or adapted to the student's course level and topic (chemistry or biochemistry). We also cover the most important theoretical aspects of the STD experiment, calling attention to some of its limitations and drawbacks without a complex theoretical approach.

AB - Saturation transfer difference (STD) NMR has emerged as one of the most popular ligand-based NMR techniques for the study of protein ligand interactions. The success of this technique is a consequence of its robustness and the fact that it is focused on the signals of the ligand, without any need of processing NMR information about the receptor and only using small quantities of nonlabeled macromolecule. Moreover, the attractiveness of this experiment is also extendable to the classroom. In the context of a practical NMR class, this experiment is ideal to illustrate some fundamental NMR concepts, such as the nuclear Overhauser effect and relaxation in a multidisciplinary context, bridging chemistry and biochemistry with a taste of medicinal chemistry. We use the readily available human serum albumin (HSA), 6-D,L-methyl-tryptophan (6-CH3-Trp), and 7- D,L-methyl-tryptophan (7-CH3-Trp) to introduce the STD-NMR experiment and to illustrate its applicability for ligand screening, mapping of binding moieties, and determination of the dissociation constant, in a context that can be explored or adapted to the student's course level and topic (chemistry or biochemistry). We also cover the most important theoretical aspects of the STD experiment, calling attention to some of its limitations and drawbacks without a complex theoretical approach.

KW - Spectroscopy

KW - ligands

KW - high-affinity

KW - Upper-Division

KW - Interdisciplinary/Multidisciplinary

KW - Biochemistry

KW - Bioanalytical

KW - Laboratory

KW - Education/Research

KW - Instruction

KW - Learning/Manipulatives

KW - nuclear-magnetic-resonance

KW - Chemistry

KW - NMR

KW - Hands-On

KW - Proteins/Peptides

KW - spectroscopy

KW - Graduate

KW - Drugs/Pharmaceuticals

KW - Undergraduate

U2 - 10.1021/ed101169t

DO - 10.1021/ed101169t

M3 - Article

VL - 88

SP - 990

EP - 994

JO - Journal of Chemical Education

JF - Journal of Chemical Education

SN - 0021-9584

IS - 7

ER -