Probing the Intracellular Glutathione Redox Potential by In-Cell NMR Spectroscopy

被引：13

作者：

Rhieu, Steve Y. ^{[1
,5
]}

Urbas, Aaron A. ^{[1
]}

Bearden, Daniel W. ^{[2
]}

Marino, John P. ^{[3
]}

Lippa, Katrice A. ^{[4
]}

Reipa, Vytas ^{[1
]}

机构：

[1] NIST, Biosyst & Biomat Div, Gaithersburg, MD 20899 USA

[2] NIST, Div Chem Sci, Hollings Marine Lab, Charleston, SC 29412 USA

[3] NIST, Inst Biosci & Biotechnol Res, Biomol Measurement Div, Rockville, MD 20850 USA

[4] NIST, Div Chem Sci, Gaithersburg, MD 20899 USA

[5] Natl Inst Stand & Technol, Natl Res Council, Gaithersburg, MD USA

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2014年 / 53卷 / 02期

关键词：

glutathione; living cells; NMR spectroscopy; oxidative stress; redox chemistry; HYDROGEN-PEROXIDE; METABOLITES; PROTEINS; EXTRACTS; SYSTEM;

D O I：

10.1002/anie.201308004

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Non-invasive and real-time analysis of cellular redox processes has been greatly hampered by lack of suitable measurement techniques. Here we describe an in-cell nuclear magnetic resonance (NMR) based method for measuring the intracellular glutathione redox potential by direct and quantitative measurement of isotopically labeled glutathione introduced exogenously into living yeast. By using this approach, perturbations in the cellular glutathione redox homeostasis were also monitored as yeast cells were subjected to oxidative stress.

引用

页码：447 / 450

页数：4

共 20 条

[1] Response of the antioxidant Defense system to tert-butyl hydroperoxide and hydrogen peroxide in a human hepatoma cell line (HepG2) [J].

Alía, M ;

Ramos, S ;

Mateos, R ;

Bravo, L ;

Goya, L .

JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, 2005, 19 (02) :119-128

[2] Hgt1p, a high affinity glutathione transporter from the yeast Saccharomyces cerevisiae [J].

Bourbouloux, A ;

Shahi, P ;

Chakladar, A ;

Delrot, S ;

Bachhawat, AH .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (18) :13259-13265

[3] Mapping structural interactions using in-cell NMR spectroscopy (STINT-NMR) [J].

Burz, DS ;

Dutta, K ;

Cowburn, D ;

Shekhtman, A .

NATURE METHODS, 2006, 3 (02) :91-93

[4] FlgM gains structure in living cells [J].

Dedmon, MM ;

Patel, CN ;

Young, GB ;

Pielak, GJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (20) :12681-12684

[5] Glutathione and catalase provide overlapping defenses for protection against hydrogen peroxide in the yeast Saccharomyces cerevisiae [J].

Grant, CM ;

Perrone, G ;

Dawes, IW .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 253 (03) :893-898

[6] Real-time imaging of the intracellular glutathione redox potential [J].

Gutscher, Marcus ;

Pauleau, Anne-Laure ;

Marty, Laurent ;

Brach, Thorsten ;

Wabnitz, Guido H. ;

Samstag, Yvonne ;

Meyer, Andreas J. ;

Dick, Tobias P. .

NATURE METHODS, 2008, 5 (06) :553-559

[7] High-resolution multi-dimensional NMR spectroscopy of proteins in human cells [J].

Inomata, Kohsuke ;

Ohno, Ayako ;

Tochio, Hidehito ;

Isogai, Shin ;

Tenno, Takeshi ;

Nakase, Ikuhiko ;

Takeuchi, Toshihide ;

Futaki, Shiroh ;

Ito, Yutaka ;

Hiroaki, Hidekazu ;

Shirakawa, Masahiro .

NATURE, 2009, 458 (7234) :106-U11

[8] Method for determining molar concentrations of metabolites in complex solutions from two-dimensional 1H-13C NMR spectra [J].

Lewis, Ian A. ;

Schommer, Seth C. ;

Hodis, Brendan ;

Robb, Kate A. ;

Tonelli, Marco ;

Westler, William M. ;

Suissman, Michael R. ;

Markley, John L. .

ANALYTICAL CHEMISTRY, 2007, 79 (24) :9385-9390

[9] Absolute quantification of metabolites in breast cancer cell extracts by quantitative 2D 1H INADEQUATE NMR [J].

Martineau, Estelle ;

Tea, Illa ;

Akoka, Serge ;

Giraudeau, Patrick .

NMR IN BIOMEDICINE, 2012, 25 (08) :985-992

[10] Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts [J].

Millard, PJ ;

Roth, BL ;

Thi, HPT ;

Yue, ST ;

Haugland, RP .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1997, 63 (07) :2897-2905

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