Microinjected glutathione reductase crystals as indicators of the redox status in living cells.
FEBS Lett
; 447(2-3): 135-8, 1999 Mar 26.
Article
en En
| MEDLINE
| ID: mdl-10214933
The flavoenzyme glutathione reductase catalyses electron transfer reactions between two major intracellular redox buffers, namely the NADPH/NADP+ couple and the 2 glutathione/glutathione disulfide couple. On this account, microcrystals of the enzyme were tested as redox probes of intracellular compartments. For introducing protein crystals into human fibroblasts, different methods (microinjection, particle bombardment and optical tweezers) were explored and compared. When glutathione reductase crystals are present in a cytosolic environment, the transition of the yellow Eox form to the orange-red 2-electron reduced charge transfer form, EH2, is observed. Taking into account the midpoint potential of the Eox/EH2 couple, the redox potential of the cytosol was found to be < -270 mV at pH 7.4 and 37 degrees C. As a general conclusion, competent proteins in crystalline--that is signal-amplifying--form are promising probes for studying intracellular events.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Glutatión Reductasa
Límite:
Humans
Idioma:
En
Revista:
FEBS Lett
Año:
1999
Tipo del documento:
Article
País de afiliación:
Alemania
Pais de publicación:
Reino Unido