title: 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.
abstract: 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 47-48kDa 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.
authors.affiliation: Department of Biology, Defence and Agro-Forestal Biotechnology, Center of Bioproteomics, University of Basilicata, 85100 Potenza, Italy. rocco.rossano@unibas.it
authors.forename: Rocco
authors.initials: R
authors.lastname: Rossano
authors.forename: Marilena
authors.initials: M
authors.lastname: Larocca
authors.forename: Paolo
authors.initials: P
authors.lastname: Riccio
mesh_terms: D001937:Brassica / Q000201:enzymology* / Q000254:growth & development / Q000502:physiology; D002734:Chlorophyll / Q000378:metabolism; D057056:Cysteine Proteases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D035264:Flowers / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D010936:Plant Extracts / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013997:Time Factors
chemical_list: D007527:Isoenzymes; D010936:Plant Extracts; D010940:Plant Proteins; D002734:Chlorophyll; D057056:Cysteine Proteases