Abstract
The pattern of endoproteolytic activities occurring during wheat (Triticum aestivum, cultivar Chinese Spring) grain development was investigated. Total endoprotease activity, assayed in solution with azocasein as a substrate, increased during the early stages of grain development to reach a maximum at 15 d postanthesis that was maintained until the grain was mature. Endoprotease activity was also assayed in gradient polyacrylamide gels co-polymerized with gelatin. The increase in endoproteolytic activity was due to the appearance of up to 18 endoproteolytic bands that were arbitrarily classified into five groups (A, B, C, D, and E). The presence of serine, aspartic, metallo, and, to a lesser extent, thiol proteases in developing wheat grains was demonstrated by the use of class-specific protease inhibitors. The appearance of the different classes of endoproteases during seed development was subject to temporal control; serine proteases were more abundant at early stages and aspartic and metallo proteases were more abundant at later stages. At intermediate stages of development (15-20 d postanthesis), most of the endoproteases were localized in the aleurone, testa, and embryo. The content of acidic thiol proteases was low in the developing starchy endosperm.
Full Text
The Full Text of this article is available as a PDF (2.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Belozersky M. A., Dunaevsky Y. E., Voskoboynikova N. E. Isolation and properties of a metalloproteinase from buckwheat (Fagopyrum esculentum) seeds. Biochem J. 1990 Dec 15;272(3):677–682. doi: 10.1042/bj2720677. [DOI] [PMC free article] [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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Cejudo F. J., Murphy G., Chinoy C., Baulcombe D. C. A gibberellin-regulated gene from wheat with sequence homology to cathepsin B of mammalian cells. Plant J. 1992 Nov;2(6):937–948. [PubMed] [Google Scholar]
- Holwerda B. C., Rogers J. C. Purification and characterization of aleurain : a plant thiol protease functionally homologous to Mammalian cathepsin h. Plant Physiol. 1992 Jul;99(3):848–855. doi: 10.1104/pp.99.3.848. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kobrehel K., Wong J. H., Balogh A., Kiss F., Yee B. C., Buchanan B. B. Specific reduction of wheat storage proteins by thioredoxin h. Plant Physiol. 1992 Jul;99(3):919–924. doi: 10.1104/pp.99.3.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koehler S. M., Ho T. H. Hormonal regulation, processing, and secretion of cysteine proteinases in barley aleurone layers. Plant Cell. 1990 Aug;2(8):769–783. doi: 10.1105/tpc.2.8.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Macnicol P. K., Jacobsen J. V. Endosperm acidification and related metabolic changes in the developing barley grain. Plant Physiol. 1992 Mar;98(3):1098–1104. doi: 10.1104/pp.98.3.1098. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mitsuhashi W., Oaks A. Development of Endopeptidase Activities in Maize (Zea mays L.) Endosperms. Plant Physiol. 1994 Feb;104(2):401–407. doi: 10.1104/pp.104.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morita S., Fukase M., Hoshino K., Fukuda Y., Yamaguchi M., Morita Y. A serine protease in soybean seeds that acts specifically on the native alpha subunit of beta-conglycinin. Plant Cell Physiol. 1994 Oct;35(7):1049–1056. [PubMed] [Google Scholar]
- Rastogi V., Oaks A. Hydrolysis of storage proteins in barley endosperms : analysis of soluble products. Plant Physiol. 1986 Jul;81(3):901–906. doi: 10.1104/pp.81.3.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rudenskaya G. N., Bogdanova E. A., Revina L. P., Golovkin B. N., Stepanov V. M. Macluralisin--a serine proteinase from fruits of Maclura pomifera (Raf.) Schneid. Planta. 1995;196(1):174–179. doi: 10.1007/BF00193231. [DOI] [PubMed] [Google Scholar]