Assessment of glutathione/glutathione disulphide ratio and S-glutathionylated proteins in human blood, solid tissues, and cultured cells

Daniela Giustarini, Graziano Colombo, Maria Lisa Garavaglia, Emanuela Astori, Nicola Marcello Portinaro, Francesco Reggiani, Salvatore Badalamenti, Anna Maria Aloisi, Annalisa Santucci, Ranieri Rossi, Aldo Milzani, Isabella Dalle-Donne

Research output: Contribution to journalReview article

Abstract

Glutathione (GSH) is the major non-protein thiol in humans and other mammals, which is present in millimolar concentrations within cells, but at much lower concentrations in the blood plasma. GSH and GSH-related enzymes act both to prevent oxidative damage and to detoxify electrophiles. Under oxidative stress, two GSH molecules become linked by a disulphide bridge to form glutathione disulphide (GSSG). Therefore, assessment of the GSH/GSSG ratio may provide an estimation of cellular redox metabolism. Current evidence resulting from studies in human blood, solid tissues, and cultured cells suggests that GSH also plays a prominent role in protein redox regulation via S -glutathionylation, i.e., the conjugation of GSH to reactive protein cysteine residues. A number of methodologies that enable quantitative analysis of GSH/GSSG ratio and S-glutathionylated proteins (PSSG), as well as identification and visualization of PSSG in tissue sections or cultured cells are currently available. Here, we have considered the main methodologies applied for GSH, GSSG and PSSG detection in biological samples. This review paper provides an up-to-date critical overview of the application of the most relevant analytical, morphological, and proteomics approaches to detect and analyse GSH, GSSG and PSSG in mammalian samples as well as discusses their current limitations.

Original languageEnglish
Pages (from-to)360-375
Number of pages16
JournalFree Radical Biology and Medicine
Volume112
DOIs
Publication statusPublished - Nov 1 2017

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Glutathione Disulfide
Protein S
Glutathione
Cultured Cells
Blood
Cells
Tissue
Oxidation-Reduction
Mammals
Oxidative stress
Sulfhydryl Compounds
Metabolism
Disulfides
Proteomics
Cysteine
Proteins
Oxidative Stress
Visualization
Plasmas
Molecules

Keywords

  • Anti-GSH antibodies
  • Biotinylation
  • Clickable glutathione
  • Fluorescent probes
  • Glutathione
  • Proteomics
  • S-glutathionylated proteins
  • Thiol alkylation

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Assessment of glutathione/glutathione disulphide ratio and S-glutathionylated proteins in human blood, solid tissues, and cultured cells. / Giustarini, Daniela; Colombo, Graziano; Garavaglia, Maria Lisa; Astori, Emanuela; Portinaro, Nicola Marcello; Reggiani, Francesco; Badalamenti, Salvatore; Aloisi, Anna Maria; Santucci, Annalisa; Rossi, Ranieri; Milzani, Aldo; Dalle-Donne, Isabella.

In: Free Radical Biology and Medicine, Vol. 112, 01.11.2017, p. 360-375.

Research output: Contribution to journalReview article

Giustarini, Daniela ; Colombo, Graziano ; Garavaglia, Maria Lisa ; Astori, Emanuela ; Portinaro, Nicola Marcello ; Reggiani, Francesco ; Badalamenti, Salvatore ; Aloisi, Anna Maria ; Santucci, Annalisa ; Rossi, Ranieri ; Milzani, Aldo ; Dalle-Donne, Isabella. / Assessment of glutathione/glutathione disulphide ratio and S-glutathionylated proteins in human blood, solid tissues, and cultured cells. In: Free Radical Biology and Medicine. 2017 ; Vol. 112. pp. 360-375.
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AU - Giustarini, Daniela

AU - Colombo, Graziano

AU - Garavaglia, Maria Lisa

AU - Astori, Emanuela

AU - Portinaro, Nicola Marcello

AU - Reggiani, Francesco

AU - Badalamenti, Salvatore

AU - Aloisi, Anna Maria

AU - Santucci, Annalisa

AU - Rossi, Ranieri

AU - Milzani, Aldo

AU - Dalle-Donne, Isabella

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AB - Glutathione (GSH) is the major non-protein thiol in humans and other mammals, which is present in millimolar concentrations within cells, but at much lower concentrations in the blood plasma. GSH and GSH-related enzymes act both to prevent oxidative damage and to detoxify electrophiles. Under oxidative stress, two GSH molecules become linked by a disulphide bridge to form glutathione disulphide (GSSG). Therefore, assessment of the GSH/GSSG ratio may provide an estimation of cellular redox metabolism. Current evidence resulting from studies in human blood, solid tissues, and cultured cells suggests that GSH also plays a prominent role in protein redox regulation via S -glutathionylation, i.e., the conjugation of GSH to reactive protein cysteine residues. A number of methodologies that enable quantitative analysis of GSH/GSSG ratio and S-glutathionylated proteins (PSSG), as well as identification and visualization of PSSG in tissue sections or cultured cells are currently available. Here, we have considered the main methodologies applied for GSH, GSSG and PSSG detection in biological samples. This review paper provides an up-to-date critical overview of the application of the most relevant analytical, morphological, and proteomics approaches to detect and analyse GSH, GSSG and PSSG in mammalian samples as well as discusses their current limitations.

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