Physiology and Molecular Biology of Plants

, Volume 18, Issue 4, pp 337–343 | Cite as

Proteolytic activities in Phaseolus vulgaris cotyledons under copper stress

  • Inès Karmous
  • Jaouani Khadija
  • Abdelilah Chaoui
  • Ezzedine El Ferjani
Research Article
First Online:

Abstract

The changes in the protease activities of bean cotyledons were investigated in response to copper stress. Assays using synthetic substrates and specific protease inhibitors followed by activity measurements and electrophoresis analysis allowed to study the classes of enzymes involved in the storage protein mobilization during the germination of bean (Phaseolus vulgaris L) seeds, and then identify which ones were affected in the presence of 200 μM CuCl2 in the imbibition medium. Copper treatment affected embryo growth and total protease activity. The results of SDS-gelatin-PAGE show that Cu excess led to a decrease in protease activity of 45 to 66 kDa. Moreover, cysteine-, aspartic- and metallo-protease activities were markedly lowered under copper stress, while serine-protease one was enhanced as well as its activity dependent abundance in comparison with control. However, the relative distribution of major cysteine protease in H2O-germinated seeds was significantly diminished after Cu exposure. Thus, copper excess can disturb the nitrogen freeing from reserve tissues at enzymatic level; differential responses of protease classes are discussed, notably, cysteine protease in the way of storage protein mobilization and serine protease in protective mechanism one.

Keywords

Bean Copper Cotyledon Endoprotease Germination 

Abbreviations

ACA

Azocaseinolytic activity

AP

Aspartic protease

CP

Cysteine protease

DAN

Dicyclohexylaminenitrite

E-64

L-trans-epoxysuccinyl-leucylamide-4-guanidino-butane

MP

Metalloprotease

NEM

N-ethylmaleimide

PMSF

Phenylmethylsulfonylfluoride

SP

Serine protease

STI

Soybean trypsin inhibitor

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Notes

Acknowledgements

Scientific work was supported by the Tunisian Ministry of Higher Education and Scientific Research.

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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2012

Authors and Affiliations

  • Inès Karmous
    • 1
  • Jaouani Khadija
    • 1
  • Abdelilah Chaoui
    • 1
  • Ezzedine El Ferjani
    • 1
  1. 1.Département des Sciences de la Vie, Faculté des Sciences de BizerteBio-Physiologie CellulairesZarzounaTunisie

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