Dinitrophenyl glutathione efflux from human erythrocytes is primary active ATP-dependent transport

E. F. LaBelle, S. V. Singh, Satish Srivastava, Y. C. Awasthi

Research output: Contribution to journalArticle

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Abstract

Dinitrophenyl S-glutathione is accumulated by inside-out vesicles made from human erythrocytes in a process totally dependent on ATP and Mg2+. The vesicles were shown to accumulate dinitrophenyl S-glutathione against a concentration gradient. The vesicles were able to concentrate this glutathione derivative even in the absence of membrane potential. This indicated that the ATP-dependent uptake of dinitrophenyl S-glutathione by inside-out vesicles represented an active transport process. Neither extravesicular EGTA nor intravesicular ouabain inhibited the transport process, indicating that neither the Ca2+-ATPase nor the Na+,K+-ATPase were involved. These results indicated that dinitrophenyl S-glutathione uptake by inside-out vesicles probably represented primary active transport. The uptake of dinitrophenyl S-glutathione was a linear function of time (up to 5 h) and vesicle protein. The rate of uptake was optimal between pH 7.0 and 8.0 and at 37°C. The K(m) values determined for dinitrophenyl S-glutathione and ATP were 0.29 mM and 1 mM, respectively. The transport process was completely inhibited by vanadate and by p-hydroxymercuribenzene sulphonate and inhibited to a lesser extent by N-ethylmaleimide. GTP could efficiently substitute for ATP as an energy source for the transport process, but CTP and UTP were comparatively much less effective.

Original languageEnglish (US)
Pages (from-to)443-449
Number of pages7
JournalBiochemical Journal
Volume238
Issue number2
StatePublished - 1986

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Glutathione
Adenosine Triphosphate
Erythrocytes
Active Biological Transport
Cytidine Triphosphate
Uridine Triphosphate
Ethylmaleimide
Vanadates
Calcium-Transporting ATPases
Egtazic Acid
Ouabain
Guanosine Triphosphate
Membrane Potentials
Adenosine Triphosphatases
Derivatives
Membranes
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

LaBelle, E. F., Singh, S. V., Srivastava, S., & Awasthi, Y. C. (1986). Dinitrophenyl glutathione efflux from human erythrocytes is primary active ATP-dependent transport. Biochemical Journal, 238(2), 443-449.

Dinitrophenyl glutathione efflux from human erythrocytes is primary active ATP-dependent transport. / LaBelle, E. F.; Singh, S. V.; Srivastava, Satish; Awasthi, Y. C.

In: Biochemical Journal, Vol. 238, No. 2, 1986, p. 443-449.

Research output: Contribution to journalArticle

LaBelle, EF, Singh, SV, Srivastava, S & Awasthi, YC 1986, 'Dinitrophenyl glutathione efflux from human erythrocytes is primary active ATP-dependent transport', Biochemical Journal, vol. 238, no. 2, pp. 443-449.
LaBelle, E. F. ; Singh, S. V. ; Srivastava, Satish ; Awasthi, Y. C. / Dinitrophenyl glutathione efflux from human erythrocytes is primary active ATP-dependent transport. In: Biochemical Journal. 1986 ; Vol. 238, No. 2. pp. 443-449.
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