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. 1990 Jun;93(2):748–757. doi: 10.1104/pp.93.2.748

Calcium Transport in the Green Alga Mesotaenium caldariorum1

Preliminary Characterization and Subcellular Distribution

Tom Berkelman 1, J Clark Lagarias 1
PMCID: PMC1062579  PMID: 16667532

Abstract

The subcellular localization and biochemical characterization of calcium transport were studied in the unicellular green alga Mesotaenium caldariorum. Membrane fractions prepared by osmotic lysis of Mesotaenium protoplasts exhibit high rates of ATP-dependent calcium uptake. Sucrose gradient centrifugation separates two pools of activity, which display specific activities for calcium transport as high as 15 nanomoles Ca2+ per minute per milligram of protein. Marker enzyme analysis shows that this dual distribution of calcium transport activity is similar to that of vanadate-insensitive ATPase and pyrophosphatase, activities considered to be associated with the tonoplast. Plasma membranes, endoplasmic reticulum vesicles, mitochondrial membranes, and thylakoids band at higher densities than either calcium transport fraction. Both pools of ATP-dependent calcium uptake contain two components which are not separable on sucrose gradients but can be distinguished on the basis of inhibitor sensitivity. One component is inhibited by nigericin or trimethyltin chloride (I50 values of 3 nanomolar and 4 micromolar, respectively), while the other component is vanadate sensitive (I50 of 25 micromolar). These results suggest that direct Ca2+ transport and Ca2+/H+ antiport activities are present in both sucrose gradient fractions.

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Selected References

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  1. Buckhout T. J. Characterization of Ca Transport in Purified Endoplasmic Reticulum Membrane Vesicles from Lepidium sativum L. Roots. Plant Physiol. 1984 Dec;76(4):962–967. doi: 10.1104/pp.76.4.962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bush D. R., Sze H. Calcium transport in tonoplast and endoplasmic reticulum vesicles isolated from cultured carrot cells. Plant Physiol. 1986 Feb;80(2):549–555. doi: 10.1104/pp.80.2.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cooper T. G., Beevers H. Mitochondria and glyoxysomes from castor bean endosperm. Enzyme constitutents and catalytic capacity. J Biol Chem. 1969 Jul 10;244(13):3507–3513. [PubMed] [Google Scholar]
  4. Gallagher S. R., Leonard R. T. Effect of vanadate, molybdate, and azide on membrane-associated ATPase and soluble phosphatase activities of corn roots. Plant Physiol. 1982 Nov;70(5):1335–1340. doi: 10.1104/pp.70.5.1335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Giannini J. L., Ruiz-Cristin J., Briskin D. P. Calcium Transport in Sealed Vesicles from Red Beet (Beta vulgaris L.) Storage Tissue : II. Characterization of Ca Uptake into Plasma Membrane Vesicles. Plant Physiol. 1987 Dec;85(4):1137–1142. doi: 10.1104/pp.85.4.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gross J., Marmé D. ATP-dependent Ca uptake into plant membrane vesicles. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1232–1236. doi: 10.1073/pnas.75.3.1232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hinson D. L., Webber R. J. Miniaturization of the BCA protein assay. Biotechniques. 1988 Jan;6(1):14–19. [PubMed] [Google Scholar]
  9. Hodges T. K., Leonard R. T. Purification of a plasma membrane-bound adenosine triphosphatase from plant roots. Methods Enzymol. 1974;32:392–406. doi: 10.1016/0076-6879(74)32039-3. [DOI] [PubMed] [Google Scholar]
  10. Kidd D. G., Lagarias J. C. Phytochrome from the green alga Mesotaenium caldariorum. Purification and preliminary characterization. J Biol Chem. 1990 Apr 25;265(12):7029–7035. [PubMed] [Google Scholar]
  11. Lew R. R., Briskin D. P., Wyse R. E. Ca uptake by endoplasmic reticulum from zucchini hypocotyls : the use of chlorotetracycline as a probe for ca uptake. Plant Physiol. 1986 Sep;82(1):47–53. doi: 10.1104/pp.82.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marin B., Gidrol X., Chrestin H., D'Auzac J. The tonoplast proton-translocating ATPase of higher plants as a third class of proton-pumps. Biochimie. 1986 Dec;68(12):1263–1277. doi: 10.1016/s0300-9084(86)80078-5. [DOI] [PubMed] [Google Scholar]
  13. Meissner G. Isolation and characterization of two types of sarcoplasmic reticulum vesicles. Biochim Biophys Acta. 1975 Apr 21;389(1):51–68. doi: 10.1016/0005-2736(75)90385-5. [DOI] [PubMed] [Google Scholar]
  14. PENNINGTON R. J. Biochemistry of dystrophic muscle. Mitochondrial succinate-tetrazolium reductase and adenosine triphosphatase. Biochem J. 1961 Sep;80:649–654. doi: 10.1042/bj0800649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SAGER R., GRANICK S. Nutritional studies with Chlamydomonas reinhardi. Ann N Y Acad Sci. 1953 Oct 14;56(5):831–838. doi: 10.1111/j.1749-6632.1953.tb30261.x. [DOI] [PubMed] [Google Scholar]
  16. Schumaker K. S., Sze H. A Ca/H Antiport System Driven by the Proton Electrochemical Gradient of a Tonoplast H-ATPase from Oat Roots. Plant Physiol. 1985 Dec;79(4):1111–1117. doi: 10.1104/pp.79.4.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Serlin B. S., Roux S. J. Modulation of chloroplast movement in the green alga Mougeotia by the Ca2+ ionophore A23187 and by calmodulin antagonists. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6368–6372. doi: 10.1073/pnas.81.20.6368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  19. TAUSSKY H. H., SHORR E. A microcolorimetric method for the determination of inorganic phosphorus. J Biol Chem. 1953 Jun;202(2):675–685. [PubMed] [Google Scholar]
  20. Takeshige K., Tazawa M., Hager A. Characterization of the H Translocating Adenosine Triphosphatase and Pyrophosphatase of Vacuolar Membranes Isolated by Means of a Perfusion Technique from Chara corallina. Plant Physiol. 1988 Apr;86(4):1168–1173. doi: 10.1104/pp.86.4.1168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tolbert N. E. Isolation of subcellular organelles of metabolism on isopycnic sucrose gradients. Methods Enzymol. 1974;31:734–746. doi: 10.1016/0076-6879(74)31077-4. [DOI] [PubMed] [Google Scholar]
  22. Widholm J. M. The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol. 1972 Jul;47(4):189–194. doi: 10.3109/10520297209116483. [DOI] [PubMed] [Google Scholar]
  23. Williamson R. E., Ashley C. C. Free Ca2+ and cytoplasmic streaming in the alga Chara. Nature. 1982 Apr 15;296(5858):647–650. doi: 10.1038/296647a0. [DOI] [PubMed] [Google Scholar]

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