title: POB1 over-expression inhibits RLIP76-mediated transport of glutathione-conjugates, drugs and promotes apoptosis.
abstract: RLIP76 (RALBP1) is a Ral-binding nucleotidase which functions as an energy-dependent transporter for glutathione (GSH)-conjugates as well as structurally unrelated xenobiotics. Partner of RALBP1 (POB1), also referred to as REPS2, was identified as the human RLIP76-binding protein, which contains a coiled-coil C-terminal region that binds with the RLIP76. Recent studies show that over-expression of POB1 in prostate cancer cells induces apoptosis. In present studies, we have purified POB1 and one of its deletion mutants POB1(1-512) (lacking the RLIP76-binding domain), and examined their effect on the transport activity of RLIP76. Both doxorubicin and a model GSH-conjugate, dinitrophenyl-S-glutathione (DNP-SG), transport were inhibited by POB1 in a concentration-dependent manner but not by POB1(1-512), lacking RLIP76-binding site. Liposomal delivery of recombinant POB1 to H358 (NSCLC) cancer cells caused apoptosis in a concentration-dependent manner, whereas the POB1 mutant deficient in RLIP76-binding site did not exert this effect. Augmentation of cellular POB1 resulted in increased intracellular DOX-accumulation as well as decreased rate of efflux from cells. These results show for the first time that POB1 can regulate the transport function of RLIP76 and are consistent with our previous studies showing that inhibition of RLIP76 induces apoptosis in cancer cells through the accumulation of endogenously formed GSH-conjugates.
authors.affiliation: Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX 76019-0065, USA.
authors.forename: Sushma
authors.initials: S
authors.lastname: Yadav
authors.forename: Ewa
authors.initials: E
authors.lastname: Zajac
authors.forename: Sharad S
authors.initials: SS
authors.lastname: Singhal
authors.forename: Jyotsana
authors.initials: J
authors.lastname: Singhal
authors.forename: Kenneth
authors.initials: K
authors.lastname: Drake
authors.forename: Yogesh C
authors.initials: YC
authors.lastname: Awasthi
authors.forename: Sanjay
authors.initials: S
authors.lastname: Awasthi
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000008:administration & dosage; D017209:Apoptosis / Q000187:drug effects*; D001692:Biological Transport / Q000187:drug effects; D002135:Calcium-Binding Proteins; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000008:administration & dosage*; D004351:Drug Resistance / Q000187:drug effects; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000008:administration & dosage* / Q000378:metabolism; D008297:Male; D011471:Prostatic Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D002135:Calcium-Binding Proteins; D020690:GTPase-Activating Proteins; D047908:Intracellular Signaling Peptides and Proteins; C095637:RALBP1 protein, human; C489722:REPS2 protein, human; D004317:Doxorubicin; D005978:Glutathione