Summary
Storage protein bodies from sunflower cotyledons during early stages of seed germination were isolated on sucrose density gradients by isopycnic centrifugation. The density of this organelle on the gradients ranged between 1.26 and 1.36 g cm-3. A proteinase with a pH optimum of 5.2 was associated with this organelle, and is probably responsible for degradation of storage protein. A NADH-dependent cytochrome-c reductase, a membrane marker enzyme with a pH optimum of 8.4, was also present in this organelle fraction.
Similar content being viewed by others
Abbreviations
- LPA:
-
for l-lysine-p-nitroanilide
- LPAase:
-
for the peptidase which hydrolyzes this peptide
References
Arnon, D. I.: Copper enzyme in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15 (1949).
Beevers, L.: Protein degradation and proteolytic activity in the cotyledons of germinating peas seeds (Pisum sativum). Phytochem. 7, 1837–1844 (1968).
Beevers, L., Spittstoesser, E. W.: Protein and nucleic acids metabolism in germinating peas. J. exp. Bot. 19, 698–711 (1968).
Briarty, L. G., Coult, D. A., Boulter, D.: Protein bodies in germinating seeds of Vicia faba. J. exp. Bot. 21, 513–524 (1970).
Donaldson, R. P.: Biochemical studies on the membranes of peroxisomes and glyoxysomes. Doct. dissert., Michigan State University, East Lansing (1971).
Donaldson, R. P., Schnarrenberger, C., Tolbert, N. E.: Cytochrome c reductase of microbodies, mitochondria, and microbodies from plants and rat liver. (Abstr.) Plant Physiol. 28, 1–47 (1971).
Erlanger, B. F., Kokowsky, N., Cohen, W.: The preparation and properties of two new chromogenic substrates of trypsin. Arch. Biochem. 95, 271–278 (1961).
Gray, R. W., Arsenis, C., Jeffay, H.: Neutral protease activity associated with the rat liver peroxisomal fraction. Biochem. biophys. Acta (Amst.) 222, 627–636 (1970).
Häcker, M.: Der Abbau von Speicherprotein und die Bildung von Plastiden in den Kotyledonen des Senfkeimlings (Sinapis alba L.) unter dem Einfluß des Phytochroms. Planta (Berl.) 76, 309–325 (1967).
Jacobson, J. V., Varner, J. E.: Gibberellic acid-induced synthesis of protease by isolated aleurone layers of barley. Plant Physiol. 42, 1596–1600 (1967).
Lowry, O. H., Rosebrough, N. J., Farr, L. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951).
Lück, H.: Catalase. In: Methods of enzymatic analysis (H. U. Bergmeyer, ed.), p. 885–894. New York: Acad. Press 1965.
Martin, E. M., Morton, R. K.: Enzymic properties of microsomes and mitochondria from Silver Beet. Biochem. J. 62, 696–704 (1956).
Morton, R. K., Raison, J. K.: The separate incorporation of amino acids into storage and soluble proteins catalysed by two independent systems isolated from developing wheat endosperm. Biochem. J. 91, 528–539 (1964).
Opik, H.: Changes in cell fine structure in the cotyledons of Phaseolus vulgaris L. during germination. J. exp. Bot. 17, 427–439 (1966).
Ory, R. L., Henningsen, K. W.: Enzymes associated with protein bodies isolated from ungerminated barley seeds. Plant Physiol. 44, 1488–1498 (1969).
Randall, D. D., Tolbert, N. E.: 3-Phosphoglycerate phosphatase. III. Activity associated with starch particles. Plant Physiol. 48, 488–492 (1971).
Schnarrenberger, C., Oeser, A., Tolbert, N. E.: Development of microbodies in sunflower cotyledons and castor bean endosperm during germination. Plant. Physiol. 48, 566–574 (1971).
Shain, Y., Mayer, A. M.: Proteolytic enzymes and endogenous trypsin inhibitor in germinating lettuce seeds. Physiol. Plantarum (Copenh.) 18, 853–859 (1965).
Smith, D. L., Flinn, A. M.: Histology and histochemistry of the cotyledons of Pisum arvense L. during germination. Planta (Berl.) 74, 72–85 (1967).
Soedigo, R., Gruber, M.: Purification and some properties of a protease from pea seeds, Pisum sativum L., spp. Arvense A and G. Biochim. biophys. Acta (Amst.) 44, 315–323 (1960).
Tolbert, N. E., Oesser, A., Kisaki, T., Hageman, R. H., Yamazaki, R. K.: Peroxisomes from spinach leaves containing enzymes related to glycolate metabolism. J. biol. Chem. 243, 5479–5184 (1968).
Tombs, M. P.: Protein bodies of the soybean. Plant Physiol. 42, 797–813 (1967).
Tronier, B., Ory, R. L., Henningsen, K. W.: Characterization of the fine structure and proteins from barley protein bodies. Phytochem. 10, 1207–1211 (1971).
Varner, J. E., Schidlovsky, G.: Intracellular distribution of proteins in pea cotyledons. Plant Physiol. 38, 139–144 (1963).
Wiley, L., Ashton, F. M.: Influence of the embryonic axis on protein hydrolysis in codyledons of Cucurbita maxima. Physiol. Plantarum (Copenh.) 20, 688–696 (1967).
Yemm, E. W., Cocking, E. C.: The determination of amino acids with ninhydrin. Analyst 80, 209–213 (1955).
Young, J. L., Varner, J. E.: Enzyme synthesis in the cotyledons of germinating seeds. Arch. Biochem. 84, 71–78 (1959).
Author information
Authors and Affiliations
Additional information
This work was supported in part by the National Science Foundation Grant GB-17543, and published as Journal Article No. 5736 of the Michigan Agricultural Experiment Station.
Supported by a Deutsche Forschungsgemeinschaft Fellowship.
Rights and permissions
About this article
Cite this article
Schnarrenberger, C., Oeser, A. & Tolbert, N.E. Isolation of protein bodies on sucrose gradients. Planta 104, 185–194 (1972). https://doi.org/10.1007/BF00387073
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00387073