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

• Authors: Rajendra Sharma , Abha Sharma , Yusong Yang , Sanjay Awasthi , Sharad S Singhal , Piotr Zimniak , Yogesh C Awasthi
• Languages of publication: EN

Abstracts

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 μM, 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.

Keywords

EN

RLIP76; 4-hydroxynonenal; Ral-binding GTPase activating protein; transport; glutathione conjugate of 4-hydroxynonenal

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2002
• Volume: 49
• Issue: 3
• Pages: 693-701

Dates

published

2002

received

2002-08-27

accepted

2002-09-04

Contributors

author

Rajendra Sharma

• Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX, U.S.A.

author

Abha Sharma

• Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX, U.S.A.

author

Yusong Yang

• Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX, U.S.A.

author

Sanjay Awasthi

• Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX, U.S.A.

author

Sharad S Singhal

• Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX, U.S.A.

author

Piotr Zimniak

• Department of Internal Medicine and Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, and Central Arkansas Veterans Healthcare System, Little Rock, AR, U.S.A

author

Yogesh C Awasthi

• Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX, U.S.A.

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