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Journal of Experimental Botany, Vol 50, 1743-1750, Copyright © 1999 by Oxford University Press

ARTICLES

Cytosolic and nuclear localization of the cytolytic and insecticidal plant protein enterolobin

C Lima, M Grossi de Sa, D Kalume, P Roepstorff, L Morhy, C Ricart and M Sousa
Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, University of Brasilia, Brasilia, DF, 70.910-900, Brazil; Laboratory of Molecular Biology, Cenargen-Embrapa, Brasilia, DF, Brazil; Protein Research Group, Department of Molecular Biology, Odense University, Odense, Denmark; Corresponding author; Fax: +55 61 272 4548; E-mail: mvsousa@unb.br

Enterolobin is a 52 905 Da cytolytic, inflammatory and insecticidal protein obtained from seeds of the Brazilian tree Enterolobium contortisiliquum. It shares structural and functional similarities with aerolysins, pore-forming toxins from Aeromonas bacteria. These characteristics suggest that enterolobin might play a defence role in the plant. In order to determine the localization and distribution of this phytocytolysin in the seed cells, immunocytochemical labelling experiments were carried out using rabbit anti-enterolobin serum and colloidal gold conjugated with goat anti-rabbit IgG. Light microscopy results indicated that labelling was widely distributed in the cytosol (33%) of cotyledon cells and in the nucleus (67%). Electron microscopy results confirmed the presence of enterolobin in both the cytosol and nucleus. Putative nuclear localization signals (NLS) were found in the sequence of enterolobin.Keywords: Enterolobium contortisiliquum, plant cytolysin, cytolytic protein, immunocytolocalization, nuclear localization signal
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