Functional reconstitution of Ral-binding GTPase activating protein, RLIP76, in proteoliposomes catalyzing ATP-dependent transport of glutathione conjugate of 4-hydroxynonenal

• Autorzy: Sharma R. , Sharma A. , Yang Y. , Awasthi S. , Singhal S.S. , Zimniak P. , Awasthi Y.C.
• Języki publikacji: EN

Abstrakty

EN

Earlier studies from our laboratories have shown that RLIP76, a previously de­scribed Ral-binding GTPase activating protein (Jullien-Flores et al., 1995, J. Biol. Chem. 270: 22473), is identical with the xenobiotic transporter DNP-SG ATPase, and can catalyze ATP-dependent transport of glutathione-conjugates as well as doxorubin (Awasthi et al., 2000, Biochemistry, 39: 9327). We have now reconstituted purified bacterially expressed RLIP76 in proteoliposomes, and have studied ATP-dependent uptake of the glutathione conjugate of 4-hydroxynonenal (GS-HNE) by these vesicles. Results of these studies show that RLIP76 reconstituted in proteoliposomes catalyzes ATP-dependent transport of GS-HNE against a concentration gradient. The transport of GS-HNE is saturable with respect to ATP as well as GS-HNE with Km values of 1.4 mM and 2.5 ,«M, respectively. These studies demonstrate that RLIP76 mediates active transport of GS-HNE, and are consistent with our previous work showing that RLIP76-mediated efflux of GS-HNE regulates the intracellular concentration of 4-HNE and thereby affects 4-HNE mediated signaling.

Słowa kluczowe

EN

transport; GTPase activating protein; proteoliposome; ATPase; glutathione; glutathione conjugation; xenobiotic transporter; 4-hydroxynonenal

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2002
• Tom: 49
• Numer: 3
• Opis fizyczny: p.693-701,fig.

Twórcy

autor

Sharma R.

• University of Texas Medical Branch, Galveston, TX 77555-0647, USA

autor

Sharma A.

autor

Yang Y.

autor

Awasthi S.

autor

Singhal S.S.

autor

Zimniak P.

autor

Awasthi Y.C.

Bibliografia

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