Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1995 Apr;107(4):1257–1268. doi: 10.1104/pp.107.4.1257

Magnesium Adenosine 5[prime]-Triphosphate-Energized Transport of Glutathione-S-Conjugates by Plant Vacuolar Membrane Vesicles.

Z S Li 1, Y Zhao 1, P A Rea 1
PMCID: PMC157260  PMID: 12228432

Abstract

By characterization of the uptake of glutathione-S-conjugates, principally dinitrophenyl-S-glutathione (DNP-GS), by vacuolar membrane vesicles, we demonstrate that a subset of energy-dependent transport processes in plants are not H+-coupled but instead are directly energized by MgATP. The most salient features of this transport pathway are: (a) its specific, obligate requirement for MgATP as energy source; (b) the necessity for hydrolysis of the [gamma]-phosphate of MgATP for uptake; (c) the insensitivity of uptake to uncouplers of the transtonoplast H+ gradient (carbonylcyanide 4-trifluoromethoxyphenylhydrazone, gramicidin-D, and NH4Cl); (d) its pronounced sensitivity to vanadate and partial inhibition by vinblastine and verapamil; (e) the lack of chemical modification of DNP-GS either during or after transport; (f) the capacity of S-conjugates of chloroacetanilide herbicides, such as metolachlor-GS, but not free herbicide, to inhibit uptake; and (g) the ability of vacuolar membrane vesicles purified from a broad range of plant species, including Arabidopsis, Beta, Vigna, and Zea, to mediate MgATP-dependent, H+-electrochemical potential difference-independent DNP-GS uptake. On the basis of these findings it is proposed that the transport of DNP-GS across the vacuolar membrane of plant cells is catalyzed by a glutathione-conjugate transporter that directly employs MgATP rather than the energy contained in the transtonoplast H+-electrochemical potential difference to drive uptake. The broad distribution of the vacuolar DNP-GS transporter and its inhibition by metolachlor-GS are consistent with the notion that it plays a general role in the vacuolar sequestration of glutathione-conjugable cytotoxic agents.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akerboom T. P., Narayanaswami V., Kunst M., Sies H. ATP-dependent S-(2,4-dinitrophenyl)glutathione transport in canalicular plasma membrane vesicles from rat liver. J Biol Chem. 1991 Jul 15;266(20):13147–13152. [PubMed] [Google Scholar]
  2. Ames G. F. Bacterial periplasmic transport systems: structure, mechanism, and evolution. Annu Rev Biochem. 1986;55:397–425. doi: 10.1146/annurev.bi.55.070186.002145. [DOI] [PubMed] [Google Scholar]
  3. Brockmöller J., Gross D., Kerb R., Drakoulis N., Roots I. Correlation between trans-stilbene oxide-glutathione conjugation activity and the deletion mutation in the glutathione S-transferase class mu gene detected by polymerase chain reaction. Biochem Pharmacol. 1992 Feb 4;43(3):647–650. doi: 10.1016/0006-2952(92)90591-6. [DOI] [PubMed] [Google Scholar]
  4. Cornwell M. M., Tsuruo T., Gottesman M. M., Pastan I. ATP-binding properties of P glycoprotein from multidrug-resistant KB cells. FASEB J. 1987 Jul;1(1):51–54. doi: 10.1096/fasebj.1.1.2886389. [DOI] [PubMed] [Google Scholar]
  5. Doige C. A., Yu X., Sharom F. J. ATPase activity of partially purified P-glycoprotein from multidrug-resistant Chinese hamster ovary cells. Biochim Biophys Acta. 1992 Aug 24;1109(2):149–160. doi: 10.1016/0005-2736(92)90078-z. [DOI] [PubMed] [Google Scholar]
  6. Foote S. J., Thompson J. K., Cowman A. F., Kemp D. J. Amplification of the multidrug resistance gene in some chloroquine-resistant isolates of P. falciparum. Cell. 1989 Jun 16;57(6):921–930. doi: 10.1016/0092-8674(89)90330-9. [DOI] [PubMed] [Google Scholar]
  7. Habig W. H., Pabst M. J., Jakoby W. B. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974 Nov 25;249(22):7130–7139. [PubMed] [Google Scholar]
  8. Higgins C. F. ABC transporters: from microorganisms to man. Annu Rev Cell Biol. 1992;8:67–113. doi: 10.1146/annurev.cb.08.110192.000435. [DOI] [PubMed] [Google Scholar]
  9. Ishikawa T., Müller M., Klünemann C., Schaub T., Keppler D. ATP-dependent primary active transport of cysteinyl leukotrienes across liver canalicular membrane. Role of the ATP-dependent transport system for glutathione S-conjugates. J Biol Chem. 1990 Nov 5;265(31):19279–19286. [PubMed] [Google Scholar]
  10. Juranka P. F., Zastawny R. L., Ling V. P-glycoprotein: multidrug-resistance and a superfamily of membrane-associated transport proteins. FASEB J. 1989 Dec;3(14):2583–2592. doi: 10.1096/fasebj.3.14.2574119. [DOI] [PubMed] [Google Scholar]
  11. Kerem B., Rommens J. M., Buchanan J. A., Markiewicz D., Cox T. K., Chakravarti A., Buchwald M., Tsui L. C. Identification of the cystic fibrosis gene: genetic analysis. Science. 1989 Sep 8;245(4922):1073–1080. doi: 10.1126/science.2570460. [DOI] [PubMed] [Google Scholar]
  12. Kitamura T., Jansen P., Hardenbrook C., Kamimoto Y., Gatmaitan Z., Arias I. M. Defective ATP-dependent bile canalicular transport of organic anions in mutant (TR-) rats with conjugated hyperbilirubinemia. Proc Natl Acad Sci U S A. 1990 May;87(9):3557–3561. doi: 10.1073/pnas.87.9.3557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kobayashi K., Sogame Y., Hayashi K., Nicotera P., Orrenius S. ATP stimulates the uptake of S-dinitrophenylglutathione by rat liver plasma membrane vesicles. FEBS Lett. 1988 Nov 21;240(1-2):55–58. doi: 10.1016/0014-5793(88)80339-9. [DOI] [PubMed] [Google Scholar]
  14. Kuchler K., Sterne R. E., Thorner J. Saccharomyces cerevisiae STE6 gene product: a novel pathway for protein export in eukaryotic cells. EMBO J. 1989 Dec 20;8(13):3973–3984. doi: 10.1002/j.1460-2075.1989.tb08580.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kunst M., Sies H., Akerboom T. P. ATP-stimulated uptake of S-(2,4-dinitrophenyl)glutathione by plasma membrane vesicles from rat liver. Biochim Biophys Acta. 1989 Jul 24;983(1):123–125. doi: 10.1016/0005-2736(89)90389-1. [DOI] [PubMed] [Google Scholar]
  16. Maeshima M., Yoshida S. Purification and properties of vacuolar membrane proton-translocating inorganic pyrophosphatase from mung bean. J Biol Chem. 1989 Nov 25;264(33):20068–20073. [PubMed] [Google Scholar]
  17. McGrath J. P., Varshavsky A. The yeast STE6 gene encodes a homologue of the mammalian multidrug resistance P-glycoprotein. Nature. 1989 Aug 3;340(6232):400–404. doi: 10.1038/340400a0. [DOI] [PubMed] [Google Scholar]
  18. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  19. Rea P. A., Britten C. J., Sarafian V. Common identity of substrate binding subunit of vacuolar h-translocating inorganic pyrophosphatase of higher plant cells. Plant Physiol. 1992 Oct;100(2):723–732. doi: 10.1104/pp.100.2.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Riordan J. R., Rommens J. M., Kerem B., Alon N., Rozmahel R., Grzelczak Z., Zielenski J., Lok S., Plavsic N., Chou J. L. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science. 1989 Sep 8;245(4922):1066–1073. doi: 10.1126/science.2475911. [DOI] [PubMed] [Google Scholar]
  21. Safa A. R., Glover C. J., Sewell J. L., Meyers M. B., Biedler J. L., Felsted R. L. Identification of the multidrug resistance-related membrane glycoprotein as an acceptor for calcium channel blockers. J Biol Chem. 1987 Jun 5;262(16):7884–7888. [PubMed] [Google Scholar]
  22. Sze H., Ward J. M., Lai S. Vacuolar H(+)-translocating ATPases from plants: structure, function, and isoforms. J Bioenerg Biomembr. 1992 Aug;24(4):371–381. doi: 10.1007/BF00762530. [DOI] [PubMed] [Google Scholar]
  23. Valvekens D., Van Montagu M., Van Lijsebettens M. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536–5540. doi: 10.1073/pnas.85.15.5536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wilson C. M., Serrano A. E., Wasley A., Bogenschutz M. P., Shankar A. H., Wirth D. F. Amplification of a gene related to mammalian mdr genes in drug-resistant Plasmodium falciparum. Science. 1989 Jun 9;244(4909):1184–1186. doi: 10.1126/science.2658061. [DOI] [PubMed] [Google Scholar]
  25. Zhen R. G., Kim E. J., Rea P. A. Localization of cytosolically oriented maleimide-reactive domain of vacuolar H(+)-pyrophosphatase. J Biol Chem. 1994 Sep 16;269(37):23342–23350. [PubMed] [Google Scholar]
  26. Zimniak P., Awasthi Y. C. ATP-dependent transport systems for organic anions. Hepatology. 1993 Feb;17(2):330–339. [PubMed] [Google Scholar]
  27. al-Shawi M. K., Senior A. E. Characterization of the adenosine triphosphatase activity of Chinese hamster P-glycoprotein. J Biol Chem. 1993 Feb 25;268(6):4197–4206. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES