Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy

Demet Araç, Tara Murphy, Jose Rizo-Rey

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Two methods for detecting protein-protein interactions in solution using one-dimensional (1D) NMR spectroscopy are described. Both methods rely on measurement of the intensity of the strongest methyl resonance (SMR), which for most proteins is observed at 0.8-0.9 ppm. The severe resonance overlap in this region facilitates detection of the SMR at low micromolar and even sub-micromolar protein concentrations. A decreased SMR intensity in the 1H NMR spectrum of a protein mixture compared to the added SMR intensities of the isolated proteins reports that the proteins interact (SMR method). Decreased SMR intensities in 1D 13C-edited 1H NMR spectra of 13C-labeled proteins upon addition of unlabeled proteins or macromolecules also demonstrate binding (SMRC method). Analysis of the interaction between XIAP and Smac, two proteins involved in apoptosis, illustrates both methods. A study showing that phospholipids compete with the neuronal core complex for Ca2+-dependent binding to the presynaptic Ca2+-sensor synaptotagmin 1 illustrates the usefulness of the SMRC method in studying multicomponent systems.

Original languageEnglish (US)
Pages (from-to)2774-2780
Number of pages7
JournalBiochemistry
Volume42
Issue number10
DOIs
StatePublished - Mar 18 2003

Fingerprint

Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
Proteins
Synaptotagmin I
Nuclear magnetic resonance
Macromolecules
Phospholipids
Apoptosis
Sensors

ASJC Scopus subject areas

  • Biochemistry

Cite this

Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy. / Araç, Demet; Murphy, Tara; Rizo-Rey, Jose.

In: Biochemistry, Vol. 42, No. 10, 18.03.2003, p. 2774-2780.

Research output: Contribution to journalArticle

Araç, Demet ; Murphy, Tara ; Rizo-Rey, Jose. / Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy. In: Biochemistry. 2003 ; Vol. 42, No. 10. pp. 2774-2780.
@article{e97640ff9df149cdbb229a8e009b5f01,
title = "Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy",
abstract = "Two methods for detecting protein-protein interactions in solution using one-dimensional (1D) NMR spectroscopy are described. Both methods rely on measurement of the intensity of the strongest methyl resonance (SMR), which for most proteins is observed at 0.8-0.9 ppm. The severe resonance overlap in this region facilitates detection of the SMR at low micromolar and even sub-micromolar protein concentrations. A decreased SMR intensity in the 1H NMR spectrum of a protein mixture compared to the added SMR intensities of the isolated proteins reports that the proteins interact (SMR method). Decreased SMR intensities in 1D 13C-edited 1H NMR spectra of 13C-labeled proteins upon addition of unlabeled proteins or macromolecules also demonstrate binding (SMRC method). Analysis of the interaction between XIAP and Smac, two proteins involved in apoptosis, illustrates both methods. A study showing that phospholipids compete with the neuronal core complex for Ca2+-dependent binding to the presynaptic Ca2+-sensor synaptotagmin 1 illustrates the usefulness of the SMRC method in studying multicomponent systems.",
author = "Demet Ara{\cc} and Tara Murphy and Jose Rizo-Rey",
year = "2003",
month = "3",
day = "18",
doi = "10.1021/bi0272050",
language = "English (US)",
volume = "42",
pages = "2774--2780",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "10",

}

TY - JOUR

T1 - Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy

AU - Araç, Demet

AU - Murphy, Tara

AU - Rizo-Rey, Jose

PY - 2003/3/18

Y1 - 2003/3/18

N2 - Two methods for detecting protein-protein interactions in solution using one-dimensional (1D) NMR spectroscopy are described. Both methods rely on measurement of the intensity of the strongest methyl resonance (SMR), which for most proteins is observed at 0.8-0.9 ppm. The severe resonance overlap in this region facilitates detection of the SMR at low micromolar and even sub-micromolar protein concentrations. A decreased SMR intensity in the 1H NMR spectrum of a protein mixture compared to the added SMR intensities of the isolated proteins reports that the proteins interact (SMR method). Decreased SMR intensities in 1D 13C-edited 1H NMR spectra of 13C-labeled proteins upon addition of unlabeled proteins or macromolecules also demonstrate binding (SMRC method). Analysis of the interaction between XIAP and Smac, two proteins involved in apoptosis, illustrates both methods. A study showing that phospholipids compete with the neuronal core complex for Ca2+-dependent binding to the presynaptic Ca2+-sensor synaptotagmin 1 illustrates the usefulness of the SMRC method in studying multicomponent systems.

AB - Two methods for detecting protein-protein interactions in solution using one-dimensional (1D) NMR spectroscopy are described. Both methods rely on measurement of the intensity of the strongest methyl resonance (SMR), which for most proteins is observed at 0.8-0.9 ppm. The severe resonance overlap in this region facilitates detection of the SMR at low micromolar and even sub-micromolar protein concentrations. A decreased SMR intensity in the 1H NMR spectrum of a protein mixture compared to the added SMR intensities of the isolated proteins reports that the proteins interact (SMR method). Decreased SMR intensities in 1D 13C-edited 1H NMR spectra of 13C-labeled proteins upon addition of unlabeled proteins or macromolecules also demonstrate binding (SMRC method). Analysis of the interaction between XIAP and Smac, two proteins involved in apoptosis, illustrates both methods. A study showing that phospholipids compete with the neuronal core complex for Ca2+-dependent binding to the presynaptic Ca2+-sensor synaptotagmin 1 illustrates the usefulness of the SMRC method in studying multicomponent systems.

UR - http://www.scopus.com/inward/record.url?scp=0037452955&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037452955&partnerID=8YFLogxK

U2 - 10.1021/bi0272050

DO - 10.1021/bi0272050

M3 - Article

C2 - 12627942

AN - SCOPUS:0037452955

VL - 42

SP - 2774

EP - 2780

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 10

ER -