Novel function of human RLIP76: ATP-dependent transport of glutathione conjugates and doxorubicin
Abstract
Active transport of conjugated and unconjugated electrophiles out of cells is essential for cellular homeostasis. We have previously identified in human tissues a transporter, DNP-SG [S-(2, 4-dinitrophenyl)glutathione] ATPase, capable of carrying out this function [Awasthi et al. (1998) Biochemistry 37, 5231-5238, 5239-5248]. We now report the cloning of DNP-SG ATPase. The sequence of the cDNA clone was identical to that of human RLIP76, a known Ral-binding protein. RLIP76 expressed in E. coli was purified by DNP-SG affinity chromatography. Purified recombinant RLIP76: (1) had ATPase activity stimulated by DNP-SG or doxorubicin (DOX), and the K(m) values of RLIP76 for ATP, DOX, and DNP-SG were similar to those reported for DNP-SG ATPase; (2) upon reconstitution with asolectin as well as with defined lipids, catalyzed ATP-dependent transport of DNP-SG and DOX with kinetic parameters similar to those of DNP-SG ATPase; (3) when transfected into K562 cells, resulted in increased resistance to DOX, and increased ATP-dependent transport of DNP-SG and DOX by inside-out membrane vesicles from transfected cells; (4) direct uptake of purified RLIP76 protein into mammalian cells from donor proteoliposomes confers DOX resistance. These results indicate that RLIP76, in addition to its role in signal transduction, can catalyze transport of glutathione conjugates and xenobiotics, and may contribute to the multidrug resistance phenomenon.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Rlip is required for low-affinity high-capacity efflux of the glutathione conjugate of 4-HNE.
“It was later shown that Rlip is required for the low-affinity high-capacity efflux of the glutathione conjugate of 4-HNE, a genotoxic and proteotoxic peroxidation product of omega-6 polyunsaturated fatty acids (PUFAs) such as arachidonic acid and linoleic acid [11].”
From Rlip76: An Unexplored Player in Neurodegeneration and Alzheimer’s Disease? (2022) · cites this paper as [11]
4-HNE is a genotoxic and proteotoxic peroxidation product of omega-6 polyunsaturated fatty acids such as arachidonic acid and linoleic acid.
“4-HNE is a genotoxic and proteotoxic peroxidation product of omega-6 polyunsaturated fatty acids (PUFAs) such as arachidonic acid and linoleic acid [11].”
From Rlip76: An Unexplored Player in Neurodegeneration and Alzheimer’s Disease? (2022) · cites this paper as [11]
Cited by
Sorted by citations
Glutathione Homeostasis and Functions: Potential Targets for Medical Interventions
Lushchak, V. I. (2012) · Journal of Amino Acids · doi:10.1155/2012/736837 · Cited by 1,266 · see citing works
Frohnert, B. I., Long, E. K., Hahn, W. S., & Bernlohr, D. A. (2014) · Diabetes · doi:10.2337/db13-0777 · Cited by 60 · see citing works
“ONE), are glutathionylated by GSTA4, exported rapidly in an ATP-dependent manner by RLIP76, and excreted in urine (15,16).”
BackgroundInfluentialRLIP76 protein knockdown attenuates obesity due to a high-fat diet
Singhal, S. S., Figarola, J., Singhal, J., Reddy, M. A., Liu, X., Berz, D., Natarajan, R., & Awasthi, S. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.480194 · Cited by 26 · see citing works
Rlip76: An Unexplored Player in Neurodegeneration and Alzheimer’s Disease?
Hindle, A., Singh, S. P., Pradeepkiran, J. A., Bose, C., Vijayan, M., Kshirsagar, S., Sawant, N. A., & Reddy, P. H. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23116098 · Cited by 14 · see citing works
Self-citation
- Authors
- Awasthi, S., Cheng, J., Singhal, S. S., Saini, M. K., Pandya, U., Pikula, S., Bandorowicz-Pikula, J., Singh, S. V., Zimniak, P., & Awasthi, Y. C.
- Affiliations
- Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, Texas, Department of Human Biological Chemistry and Genetics, UTMB, Galveston, Texas, Nencki Institute of Experimental Biology, Warsaw, Poland, Mercy Hospital Cancer Center, Pittsburgh, Pennsylvania, and Department of Internal Medicine and Department of Biochemistry & Molecular Biology, University of Arkansas for Medical Sciences and McClellan VA Medical Center, Little Rock, Arkansas
- Instytut Biologii Doświadczalnej im. Marcelego Nenckiego (PL)
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2000-07-13
- Volume
- 39
- Issue
- 31
- Pages
- 9327-9334
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W1986301201
- Cited by
- 159
- References
- 0
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 3 · 2023: 4 · 2022: 3 · 2021: 7 · 2020: 3 · 2019: 7 · 2018: 8 · 2017: 5 · 2016: 10 · 2015: 6 · 2014: 6 · 2013: 9 · 2012: 7
- FWCI
- 1.96
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Cancer Institute (CA 77495, CA 55589, R01CA055589, R01CA077495)
- National Institute of General Medical Sciences (GM-32304, R01GM032304)
- Topics
- Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glutathione Transferases and Polymorphisms
- Keywords
- RLIP76, glutathione conjugates, doxorubicin, ATP-dependent transport, multidrug resistance, xenobiotic transport, drug efflux, K562 cells, membrane vesicles, proteoliposomes, cDNA cloning
- MeSH terms
- Adenosine Triphosphatases, Adenosine Triphosphate, Amino Acid Sequence, Amino Acids, Biological Transport, Active, Carrier Proteins, Catalysis, Cell Adhesion, Cell Membrane, Doxorubicin, Glutathione, Humans, Intracellular Fluid, Molecular Sequence Data, Peptide Fragments, Phospholipids, Proteolipids, Recombinant Proteins, Transfection, Tumor Cells, Cultured, ATP-Binding Cassette Transporters, K562 Cells, GTPase-Activating Proteins
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
