Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-27T00:15:54.407Z Has data issue: false hasContentIssue false

Purification and characterization of an aspartyl proteinase from dry jack pine seeds

Published online by Cambridge University Press:  19 September 2008

Jacqueline Bourgeois
Affiliation:
Department of Biology, Lakehead University, Thunder Bay, Ontario, Canada P7B 5E1
L. Malek*
Affiliation:
Department of Biology, Lakehead University, Thunder Bay, Ontario, Canada P7B 5E1
*
* Correspondence

Abstract

A high-molecular-weight aspartyl proteinase complex, sensitive to pepstatin A was purified to near electrophoretic homogeneity from dry seeds ofjack pine (Pinus banksiana, Lamb.). Two partial activities, endo-and exo-proteinolytic, were associated with the complex, judged by the analysis of haemoglobin hydrolysis products. The high activity of this enzyme in dry jack pine seed suggests a possible function in early germination.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1991

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anson, M.L., (1938) The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin. Journal of General Physiology 22, 7989.CrossRefGoogle ScholarPubMed
Bourgeois, J. (1989) Priming physiology and proteinase purification from resting jack pine seeds. MSc Thesis, Lakehead University, Thunder Bay, Canada.Google Scholar
Bradford, M.M. (1976) A rapid and sensitive method for the quantities of protein-dye binding. Analytical Biochemistry 72, 248254.CrossRefGoogle ScholarPubMed
Ching, T.M. (1966) Compositional changes of Douglas fir seeds during germination. Plant Physiology 41, 13131319.CrossRefGoogle ScholarPubMed
Corvol, P., Devaux, C. and Menard, J. (1973) Pepstatin, an inhibitor for renin purification by affinity chromatography. FEBS Letters 34(2),189192.CrossRefGoogle ScholarPubMed
Davis, B.J. (1964) Disc-electrophoresis. II. Method and application to human serum proteins. Annals of the New York Academy of Science 121, 404427.CrossRefGoogle ScholarPubMed
DeMartino, G.N. (1989) Purification of proteolytic enzymes. in Beynon, R.J., and Bond, J.S. (Eds) Proteolytic enzymes, a practical approach. Oxford, IRL Press.Google Scholar
Driscoll, J. and Goldberg, A.L. (1990) The proteasome (multicatalytic protease) is a component of the 1500 kDa proteolytic complex which degrades ubiquitin conjugated proteins. Journal of Biological Chemistry 265(9), 47894792.CrossRefGoogle ScholarPubMed
Gifford, D.J. (1988) An electrophoretic analysis of the seed proteins from Pinus monticola and eight other species of pine. Canadian Journal of Botany 66, 18081812.CrossRefGoogle Scholar
Goo, M. and Furusawa, K. (1955) Changes the stored food within the seeds of Pinus densiflora and P. humbergii during the course of germination by microchemical tests. Journal of the Japanese Forestry Society 37(10), 481484.Google Scholar
Hackenthal, E., Hackenthal, R. and Hilgenfendt, U. (1978) Purification and partial characterization of rat brain acid proteinase (isorenin). Biochimica et Biophysica Acta 522, 561573.CrossRefGoogle ScholarPubMed
Katsuta, M. (1961) Break-down of reserve proteinof pine seeds during germination. Journal of the Japanese Forestry Society 43(6), 241244.Google Scholar
Kay, J. (1985) Aspartic proteinases and their inhibitors. pp. 117 in Kostka, V. (Ed.) Asparticproteinases and inhibitors. Proceedings of the FEBS Advanced CourseNo.84107 Berlin, Walter de Gruyter.Google Scholar
Koehler, S. and Ho, T-H.D. (1988) Purification and characterization of gibberellic acid-induced cysteine endopeptidasesin barley aleurone layers. Plant Physiology 87, 95103.CrossRefGoogle ScholarPubMed
Kregac, I., Urh, I., Smith, R., Umezawa, H. and Turk, V. (1977) Isolation of cathepsin D by affinity chromatography on immobilized pepstatin. pp. 250254 in Turk, V. and Marks, H. (Eds) Intracellular protein catabolism II. Proceedings. New York, Plenum Press.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680685.CrossRefGoogle ScholarPubMed
Leluk, J., Otlewski, J., Wieczorek, M., Polanowski, A. and Wilusz, T. (1983) Preparation and characteristics of trypsin inhibitors from the seeds of squash (Cucurbita maxima) and zucchini (Cucurbita pepo var. giromontia) Acta Biochem. Polonica 30, 127138.Google ScholarPubMed
Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.L. (1951) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265275.CrossRefGoogle ScholarPubMed
Marciniszyn, J. Jr, Hartsack, J.A. and Tang, J. (1976) Mode of inhibition of acid proteases by pepstatin. Journal of Biological Chemistry 257(22), 70887094.CrossRefGoogle Scholar
Polanowski, A., Wilusz, T., Kolaczkowska, M.K., Wieczorek, M. and Wilimowska-Pelc, W. (1985) Purification and characterization of aspartic proteinases from Cucumis sativus and Cucurbita maxima seeds. pp. 4952 in Kostka, V. (Ed.) Aspartic proteinases and inhibitors. Proceedings of the FEBS Advanced Course No.84107. Berlin, Walter de Gruyter.CrossRefGoogle Scholar
Salmia, M.A. (1980) Inhibitors of endogenous proteinases in Scots pine seeds: fractionation and activity changes during germination. Physiologia Plantarum 48, 266270.CrossRefGoogle Scholar
Salmia, M.A. (1981a) Proteinase activities in resting and germinating seeds of scots pine; Pinus sylvestris. Physiologia Plantarum 53, 3947.CrossRefGoogle Scholar
Salmia, M.A. (1981b) Fractionation of the proteinases present in the endosperm of germinating seed of Scots pine; Pinus sylvestris. Physiologia Plantarum 51, 253258.CrossRefGoogle Scholar
Salmia, M.A. and Mikola, J.J. (1975) Activities of two peptidases in resting and germinating seeds of Scots pine; Pinus sylvestris. Physiologia Plantarum 33, 261265.CrossRefGoogle Scholar
Salmia, M.A. and Mikola, J.J. (1980) Inhibitors of endogenous proteinases in the seeds of Scots pine; Pinus sylvestris. Physiologia Plantarum 48, 126130.CrossRefGoogle Scholar
Salmia, M.A. and Mikola, J.J. (1976a) Localization and activity of carboxypeptidase in germinating seeds of Scots pine; Pinus sylvestris. Physiologia Plantarum 36, 388392.CrossRefGoogle Scholar
Salmia, M.A. and Mikola, J.J. (1976b) Localization and activity of naphthylamidases in germinating seeds of Scots pine; Pinus sylvestris. Physiologia Plantarum 38, 7377.CrossRefGoogle Scholar
Salmia, M.A., Nyman, S.A. and Mikola, J.J. (1978) Characterization of the proteinase present in germinating seeds of Scots pine; Pinus sylvestris. Physiologia Plantarum 48, 126130.CrossRefGoogle Scholar
Simola, L.K. (1974) The ultrastructure of dry and germinating seeds of Pinus sylvestris L. Acta Botanica Fennica 103, 131.Google Scholar
Swank, R.T. and Munches, K.D. (1971) Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Analytical Biochemistry 39, 462477.CrossRefGoogle ScholarPubMed
Tanaka, K. and Ichihara, A. (1990) Proteasomes (multicatalytic proteinase complexes) in eukaryotic cells. Cell Structure and Function 15, 127132.CrossRefGoogle ScholarPubMed
Wilimowska-Pelc, A., Polanowski, A., Kolaczkowska, M.K., Wieczorek, M. and Wilusz, T. (1983) Aspartyl proteinase from cucumber (Cucumis sativus) seeds.Preparation and characterization. Acta Biochimica Polonica 30, 2331.Google Scholar
Yang, J.-F. and Malek, L. (1991) Purification of a dry pea seed proteinolytic enzyme complex. Phytochemistry 39, 24872491.CrossRefGoogle Scholar