Abstract
Changing patterns of enzyme activity and solute transport in response to washing were investigated in red beet (Beta vulgaris L.) storage tissue. Washing had a pronounced effect on the plasma membrane (PM) H+-ATPase with an increase in both hydrolytic and proton-pumping activities. Immunoblotting indicated that this may be due, in part, to a higher amount of this enzyme in the PM of washed tissue. Activities of the tonoplast (V)H+-ATPase and pyrophosphatase fluctuated during a 4-d washing period, but overall showed no marked change in activity. In tissue discs sucrose (Suc), glucose (Glc), and fructose uptakes increased significantly in response to washing. Cycloheximide, cordycepin, and tunicamycin inhibited both Glc- and Suc-inducible uptake. Monensin also strongly inhibited inducible Glc uptake, but the effect on Suc was less marked. N-Ethylmaleimide inhibited both Suc and Glc uptake, with its effects being more pronounced in fresh tissue. Other protein-modifying reagents showed no significant difference in their level of inhibition between fresh and washed tissue. Transport studies, carried out using energized PM vesicles from fresh and washed tissue, indicated that there was no rise in Suc and Glc uptake rates in response to washing. Results with a range of inhibitors indicated that there was no marked change in transporter sensitivity in vesicles isolated from fresh and washed tissue. The results indicate that the well-described enhancement of solute transport in washed storage tissue may be due to an increased PM H+-ATPase activity rather than to changes in PM carrier activity or to changes in metabolism such as invertase activity.
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Selected References
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- Boutry M., Michelet B., Goffeau A. Molecular cloning of a family of plant genes encoding a protein homologous to plasma membrane H+-translocating ATPases. Biochem Biophys Res Commun. 1989 Jul 31;162(2):567–574. doi: 10.1016/0006-291x(89)92348-6. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Bush D. R. Proton-Coupled Sucrose Transport in Plasmalemma Vesicles Isolated from Sugar Beet (Beta vulgaris L. cv Great Western) Leaves. Plant Physiol. 1989 Apr;89(4):1318–1323. doi: 10.1104/pp.89.4.1318. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giannini J. L., Gildensoph L. H., Reynolds-Niesman I., Briskin D. P. Calcium Transport in Sealed Vesicles from Red Beet (Beta vulgaris L.) Storage Tissue : I. Characterization of a Ca-Pumping ATPase Associated with the Endoplasmic Reticulum. Plant Physiol. 1987 Dec;85(4):1129–1136. doi: 10.1104/pp.85.4.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giaquinta R. Evidence for Phloem loading from the apoplast: chemical modification of membrane sulfhydryl groups. Plant Physiol. 1976 Jun;57(6):872–875. doi: 10.1104/pp.57.6.872. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gronewald J. W., Cheeseman J. M., Hanson J. B. Comparison of the responses of corn root tissue to fusicoccin and washing. Plant Physiol. 1979 Feb;63(2):255–259. doi: 10.1104/pp.63.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harper J. F., Manney L., DeWitt N. D., Yoo M. H., Sussman M. R. The Arabidopsis thaliana plasma membrane H(+)-ATPase multigene family. Genomic sequence and expression of a third isoform. J Biol Chem. 1990 Aug 15;265(23):13601–13608. [PubMed] [Google Scholar]
- Katz D. B., Sussman M. R. Inhibition and Labeling of the Plant Plasma Membrane H-ATPase with N-Ethylmaleimide. Plant Physiol. 1987 Apr;83(4):977–981. doi: 10.1104/pp.83.4.977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Leigh R. A., Rees T., Fuller W. A., Banfield J. The location of acid invertase activity and sucrose in the vacuoles of storage roots of beetroot (Beta vulgaris). Biochem J. 1979 Mar 15;178(3):539–547. doi: 10.1042/bj1780539. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leonard R. T., Hanson J. B. Increased Membrane-bound Adenosine Triphosphatase Activity Accompanying Development of Enhanced Solute Uptake in Washed Corn Root Tissue. Plant Physiol. 1972 Mar;49(3):436–440. doi: 10.1104/pp.49.3.436. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Michelet B., Lukaszewicz M., Dupriez V., Boutry M. A plant plasma membrane proton-ATPase gene is regulated by development and environment and shows signs of a translational regulation. Plant Cell. 1994 Oct;6(10):1375–1389. doi: 10.1105/tpc.6.10.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mollenhauer H. H., Morré D. J., Rowe L. D. Alteration of intracellular traffic by monensin; mechanism, specificity and relationship to toxicity. Biochim Biophys Acta. 1990 May 7;1031(2):225–246. doi: 10.1016/0304-4157(90)90008-Z. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noubhani A. M., Sakr S., Denis M. H., Delrot S. Transcriptional and post-translational control of the plant plasma membrane H(+)-ATPase by mechanical treatments. Biochim Biophys Acta. 1996 Jun 11;1281(2):213–219. doi: 10.1016/0005-2736(96)00017-x. [DOI] [PubMed] [Google Scholar]
- Ohnishi T., Gall R. S., Mayer M. L. An improved assay of inorganic phosphate in the presence of extralabile phosphate compounds: application to the ATPase assay in the presence of phosphocreatine. Anal Biochem. 1975 Nov;69(1):261–267. doi: 10.1016/0003-2697(75)90585-0. [DOI] [PubMed] [Google Scholar]
- Perez C., Michelet B., Ferrant V., Bogaerts P., Boutry M. Differential expression within a three-gene subfamily encoding a plasma membrane H(+)-ATPase in Nicotiana plumbaginifolia. J Biol Chem. 1992 Jan 15;267(2):1204–1211. [PubMed] [Google Scholar]
- Sakr S., Lemoine R., Gaillard C., Delrot S. Effect of cutting on solute uptake by plasma membrane vesicles from sugar beet (Beta vulgaris L.) leaves. Plant Physiol. 1993 Sep;103(1):49–58. doi: 10.1104/pp.103.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sarafian V., Poole R. J. Purification of an h-translocating inorganic pyrophosphatase from vacuole membranes of red beet. Plant Physiol. 1989 Sep;91(1):34–38. doi: 10.1104/pp.91.1.34. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Toggenburger G., Kessler M., Semenza G. Phlorizin as a probe of the small-intestinal Na+,D-glucose cotransporter. A model. Biochim Biophys Acta. 1982 Jun 14;688(2):557–571. doi: 10.1016/0005-2736(82)90367-4. [DOI] [PubMed] [Google Scholar]
