Cross-saturation and transferred cross-saturation experiments

Takumi Ueda, Koh Takeuchi, Noritaka Nishida, Pavlos Stampoulis, Yutaka Kofuku, Masanori Osawa, Ichio Shimada

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Structural analyses of protein-protein interactions are required to reveal their functional mechanisms, and accurate protein-protein complex models, based on experimental results, are the starting points for drug development. In addition, structural information about proteins under physiologically relevant conditions is crucially important for understanding biological events. However, for proteins such as those embedded in lipid bilayers and transiently complexed with their effectors under physiological conditions, structural analyses by conventional methods are generally difficult, due to their large molecular weights and inhomogeneity. We have developed the cross-saturation (CS) method, which is an nuclear magnetic resonance measurement technique for the precise identification of the interfaces of protein-protein complexes. In addition, we have developed an extended version of the CS method, termed transferred cross-saturation (TCS), which enables the identification of the residues of protein ligands in close proximity to huge (>150kDa) and heterogeneous complexes under fast exchange conditions (>0.1s-1). Here, we discuss the outline, basic theory, and practical considerations of the CS and TCS methods. In addition, we will review the recent progress in the construction of models of protein-protein complexes, based on CS and TCS experiments, and applications of TCS to in situ analyses of biologically and medically important proteins in physiologically relevant states.

Original languageEnglish
Pages (from-to)1-45
Number of pages45
JournalQuarterly Reviews of Biophysics
Volume753
Issue number1
DOIs
Publication statusPublished - 2014 Apr 29
Externally publishedYes

Fingerprint

Proteins
Lipid Bilayers
Magnetic Resonance Spectroscopy
Molecular Weight
Ligands
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biophysics

Cite this

Ueda, T., Takeuchi, K., Nishida, N., Stampoulis, P., Kofuku, Y., Osawa, M., & Shimada, I. (2014). Cross-saturation and transferred cross-saturation experiments. Quarterly Reviews of Biophysics, 753(1), 1-45. https://doi.org/10.1017/S0033583514000043

Cross-saturation and transferred cross-saturation experiments. / Ueda, Takumi; Takeuchi, Koh; Nishida, Noritaka; Stampoulis, Pavlos; Kofuku, Yutaka; Osawa, Masanori; Shimada, Ichio.

In: Quarterly Reviews of Biophysics, Vol. 753, No. 1, 29.04.2014, p. 1-45.

Research output: Contribution to journalArticle

Ueda, T, Takeuchi, K, Nishida, N, Stampoulis, P, Kofuku, Y, Osawa, M & Shimada, I 2014, 'Cross-saturation and transferred cross-saturation experiments', Quarterly Reviews of Biophysics, vol. 753, no. 1, pp. 1-45. https://doi.org/10.1017/S0033583514000043
Ueda, Takumi ; Takeuchi, Koh ; Nishida, Noritaka ; Stampoulis, Pavlos ; Kofuku, Yutaka ; Osawa, Masanori ; Shimada, Ichio. / Cross-saturation and transferred cross-saturation experiments. In: Quarterly Reviews of Biophysics. 2014 ; Vol. 753, No. 1. pp. 1-45.
@article{01b21e6e290c4097b9dbba6a89d7900a,
title = "Cross-saturation and transferred cross-saturation experiments",
abstract = "Structural analyses of protein-protein interactions are required to reveal their functional mechanisms, and accurate protein-protein complex models, based on experimental results, are the starting points for drug development. In addition, structural information about proteins under physiologically relevant conditions is crucially important for understanding biological events. However, for proteins such as those embedded in lipid bilayers and transiently complexed with their effectors under physiological conditions, structural analyses by conventional methods are generally difficult, due to their large molecular weights and inhomogeneity. We have developed the cross-saturation (CS) method, which is an nuclear magnetic resonance measurement technique for the precise identification of the interfaces of protein-protein complexes. In addition, we have developed an extended version of the CS method, termed transferred cross-saturation (TCS), which enables the identification of the residues of protein ligands in close proximity to huge (>150kDa) and heterogeneous complexes under fast exchange conditions (>0.1s-1). Here, we discuss the outline, basic theory, and practical considerations of the CS and TCS methods. In addition, we will review the recent progress in the construction of models of protein-protein complexes, based on CS and TCS experiments, and applications of TCS to in situ analyses of biologically and medically important proteins in physiologically relevant states.",
author = "Takumi Ueda and Koh Takeuchi and Noritaka Nishida and Pavlos Stampoulis and Yutaka Kofuku and Masanori Osawa and Ichio Shimada",
year = "2014",
month = "4",
day = "29",
doi = "10.1017/S0033583514000043",
language = "English",
volume = "753",
pages = "1--45",
journal = "Quarterly Reviews of Biophysics",
issn = "0033-5835",
publisher = "Cambridge University Press",
number = "1",

}

TY - JOUR

T1 - Cross-saturation and transferred cross-saturation experiments

AU - Ueda, Takumi

AU - Takeuchi, Koh

AU - Nishida, Noritaka

AU - Stampoulis, Pavlos

AU - Kofuku, Yutaka

AU - Osawa, Masanori

AU - Shimada, Ichio

PY - 2014/4/29

Y1 - 2014/4/29

N2 - Structural analyses of protein-protein interactions are required to reveal their functional mechanisms, and accurate protein-protein complex models, based on experimental results, are the starting points for drug development. In addition, structural information about proteins under physiologically relevant conditions is crucially important for understanding biological events. However, for proteins such as those embedded in lipid bilayers and transiently complexed with their effectors under physiological conditions, structural analyses by conventional methods are generally difficult, due to their large molecular weights and inhomogeneity. We have developed the cross-saturation (CS) method, which is an nuclear magnetic resonance measurement technique for the precise identification of the interfaces of protein-protein complexes. In addition, we have developed an extended version of the CS method, termed transferred cross-saturation (TCS), which enables the identification of the residues of protein ligands in close proximity to huge (>150kDa) and heterogeneous complexes under fast exchange conditions (>0.1s-1). Here, we discuss the outline, basic theory, and practical considerations of the CS and TCS methods. In addition, we will review the recent progress in the construction of models of protein-protein complexes, based on CS and TCS experiments, and applications of TCS to in situ analyses of biologically and medically important proteins in physiologically relevant states.

AB - Structural analyses of protein-protein interactions are required to reveal their functional mechanisms, and accurate protein-protein complex models, based on experimental results, are the starting points for drug development. In addition, structural information about proteins under physiologically relevant conditions is crucially important for understanding biological events. However, for proteins such as those embedded in lipid bilayers and transiently complexed with their effectors under physiological conditions, structural analyses by conventional methods are generally difficult, due to their large molecular weights and inhomogeneity. We have developed the cross-saturation (CS) method, which is an nuclear magnetic resonance measurement technique for the precise identification of the interfaces of protein-protein complexes. In addition, we have developed an extended version of the CS method, termed transferred cross-saturation (TCS), which enables the identification of the residues of protein ligands in close proximity to huge (>150kDa) and heterogeneous complexes under fast exchange conditions (>0.1s-1). Here, we discuss the outline, basic theory, and practical considerations of the CS and TCS methods. In addition, we will review the recent progress in the construction of models of protein-protein complexes, based on CS and TCS experiments, and applications of TCS to in situ analyses of biologically and medically important proteins in physiologically relevant states.

UR - http://www.scopus.com/inward/record.url?scp=84917699524&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84917699524&partnerID=8YFLogxK

U2 - 10.1017/S0033583514000043

DO - 10.1017/S0033583514000043

M3 - Article

C2 - 24780282

AN - SCOPUS:84907598043

VL - 753

SP - 1

EP - 45

JO - Quarterly Reviews of Biophysics

JF - Quarterly Reviews of Biophysics

SN - 0033-5835

IS - 1

ER -