2-D zymographic analysis of Broccoli ( Brassica oleracea L. var. Italica) florets proteases: Follow up of cysteine protease isotypes in the course of post-harvest senescence

Zymographic analysis of Broccoli florets ( Brassica oleracea L. var. Italica) revealed the presence of acidic metallo-proteases, serine proteases and cysteine proteases. Under conditions which were denaturing for the other proteases, the study was restricted to cysteine proteases. 2-D zymography, a...

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Bibliographic Details
Published in:Journal of plant physiology 2011-09, Vol.168 (13), p.1517-1525
Main Authors: Rossano, Rocco, Larocca, Marilena, Riccio, Paolo
Format: Article
Language:English
Subjects:
Biological and medical sciences
Brassica - enzymology
Brassica - growth & development
Brassica - physiology
Broccoli
Chlorophyll - metabolism
Cysteine Proteases - analysis
Cysteine Proteases - genetics
Cysteine Proteases - metabolism
Electrophoresis, Gel, Two-Dimensional - methods
Flowers - enzymology
Flowers - growth & development
Flowers - metabolism
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
Isoelectric Focusing
Isoenzymes - analysis
Isoenzymes - genetics
Isoenzymes - metabolism
Phosphorylation
Plant Extracts - chemistry
Plant physiology and development
Plant Proteins - analysis
Plant Proteins - genetics
Plant Proteins - metabolism
Post-harvest
Proteases
Senescence
Senescence and abscission
Time Factors
Vegetative apparatus, growth and morphogenesis. Senescence
Zymography
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Summary:Zymographic analysis of Broccoli florets ( Brassica oleracea L. var. Italica) revealed the presence of acidic metallo-proteases, serine proteases and cysteine proteases. Under conditions which were denaturing for the other proteases, the study was restricted to cysteine proteases. 2-D zymography, a technique that combines IEF and zymography was used to show the presence of 11 different cysteine protease spots with molecular mass of 44 and 47–48 kDa and pIs ranging between 4.1 and 4.7. pI differences could be ascribed to different degrees of phosphorylation that partly disappeared in the presence of alkaline phosphatase. Post-harvest senescence of Broccoli florets was characterized by decrease in protein and chlorophyll contents and increase of protease activity. In particular, as determined by 2-D zymography, the presence of cysteine protease clearly increased during senescence, a finding that may represent a useful tool for the control of the aging process.
ISSN:0176-1617
1618-1328
DOI:10.1016/j.jplph.2011.02.013