The NMR signal of reduced glutathione (GSH) was monitored in intact human erythrocytes by the 1H spin-echo Carr-Purcell-Meiboom-Gill pulse sequence. Addition of GSH, which was unable to cross the erythrocyte membrane, produced an approximate two-fold increase of the GSH signal in glucose-depleted cells. Addition of oxidised glutathione (GSSG), did not affect the signal, and addition of GSH to hemolysates gave a much smaller increase. Reduction of internal GSSG by NADPH-dependent enzymes was excluded by experiments with glucose-supplied or glucose-6-phosphate dehydrogenase deficient cells. Involvement of external thiol groups of the erythrocyte membrane was shown by the lack of effect in cells treated with an impermeable thiol-blocking compound. Involvement of spectrin was indicated by the proportional loss of the effect in erythrocytes with variable genetic deficiency of spectrin. Protein-glutathione mixed disulfides appeared to be the source of the NMR response since an increase of their content, by diamide treatment or aging procedures, produced a higher GSH signal, while their reduction by permeable reductants gave the opposite effect. It is concluded that GSH can transduce its reducing power by a thiol/disulfide exchange mechanism that sequentially involves sulfur-rich proteins spanning across the erythrocyte membrane.
Ciriolo, M.R., Paci, M., Sette, M., De Martino, A., Bozzi, A., & Rotilio, G. (1993). Transduction of reducing power across the plasma membrane by reduced glutathione. A 1H-NMR spin-echo study of intact human erythrocytes. EUROPEAN JOURNAL OF BIOCHEMISTRY, 215(3), 711-718.
Tipologia: | Articolo su rivista |
Citazione: | Ciriolo, M.R., Paci, M., Sette, M., De Martino, A., Bozzi, A., & Rotilio, G. (1993). Transduction of reducing power across the plasma membrane by reduced glutathione. A 1H-NMR spin-echo study of intact human erythrocytes. EUROPEAN JOURNAL OF BIOCHEMISTRY, 215(3), 711-718. |
URL: | http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1993.tb18083.x/pdf |
IF: | Con Impact Factor ISI |
Lingua: | English |
Settore Scientifico Disciplinare: | Settore BIO/10 |
Revisione (peer review): | Sì, ma tipo non specificato |
Tipo: | Articolo |
Rilevanza: | Rilevanza internazionale |
Stato di pubblicazione: | Pubblicato |
Data di pubblicazione: | 1993 |
Titolo: | Transduction of reducing power across the plasma membrane by reduced glutathione. A 1H-NMR spin-echo study of intact human erythrocytes |
Autori: | |
Autori: | Ciriolo, MR; Paci, M; Sette, M; De Martino, A; Bozzi, A; Rotilio, G |
Appare nelle tipologie: | 01 - Articolo su rivista |