Gel electrophoretic profiles of proteinases in dark-germinated flax seeds

Jameel, Shahid ; Reddy, V. Manoranjan ; Rhodes, W. Gale ; McFadde, Bruce A. (1984) Gel electrophoretic profiles of proteinases in dark-germinated flax seeds Plant Physiology, 76 . pp. 730-734. ISSN 0032-0889

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The proteinases present in dark-germinated flax seeds (Linum usitatissimum) were studied as a function of germination at 25°C. A majority of activity was present in basic proteinases with an acidic pH optimum and a temperature optimum of 45°C in the digestion of hemoglobin. Electrophoresis in a sodium dodecyl sulfate-polyacrylamide mixture which had been polymerized with gelatin was used to separate proteins in extracts of seedlings. Subsequent activation of proteinases with Triton X-100 and resultant digestion of gelatin proved to be very reproducible and afforded detection and good quantification of various proteinase zones. An ethylenediaminetetraacetate-sensitive proteinase zone, P4 (about 60,000 daltons), appeared at day 3 after imbibition and attained maximum activity at day 4. This correlates with a rapid loss in vivo of the glyoxysomal enzyme, isocitrate lyase (EC Ethylenediaminetetraacetate also slowed the loss of isocitrate lyase activity in extracts of 4-day seedlings in a dose-dependent manner. The addition of leupeptin,α -tolylsulfonyl fluoride, Pepstatin A, p-chloromercuribenzoate, or 1,10-phenanthroline prior to, during, or after exchange of Triton X-100 for sodium dodecyl sulfate had almost no inhibitory effect upon proteinases in 4-day seedlings.

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Source:Copyright of this article belongs to American Society of Plant Biologists.
ID Code:13099
Deposited On:11 Nov 2010 06:53
Last Modified:16 May 2016 22:19

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