title: Determination of the amino acid sequence of the plant cytolysin enterolobin.
abstract: The cytolytic seed protein enterolobin from seeds of Enterolobium contortisiliquum was purified by using FPLC on a Mono Q column giving a single peak in capillary electrophoresis. The complete amino acid sequence of the plant cytolysin was determined by an automated method, yielding a molecular mass of 54,806 Da. Databank searches and sequence alignment demonstrated a high degree of sequence identity and similarity between enterolobin and bacterial aerolysins from Aeromonas hydrophila and A. sobria. Several key residues involved in oligomerization of A. hydrophila aerolysin are conserved in enterolobin. Circular dichroism measurements and structural predictions revealed that enterolobin is very rich in beta sheet, like aerolysin. Light-scattering studies revealed that enterolobin oligomerizes as a hexamer at pH levels below 7.0. NaCl concentrations above 50 mM caused dimerization of enterolobin. Dithiothreitol did not cause oligomerization.
authors.affiliation: Laboratório de Bioquímica e Química de Proteínas, Universidade de Brasília, Brasília-DF, 70910-900, Brazil.
authors.forename: W
authors.initials: W
authors.lastname: Fontes
authors.forename: M V
authors.initials: MV
authors.lastname: Sousa
authors.forename: J B
authors.initials: JB
authors.lastname: Aragão
authors.forename: L
authors.initials: L
authors.lastname: Morhy
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001427:Bacterial Toxins / Q000737:chemistry; D003603:Cytotoxins / Q000737:chemistry*; D019075:Electrophoresis, Capillary; D008027:Light; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry*; D052899:Pore Forming Cytotoxic Proteins; D011498:Protein Precursors / Q000737:chemistry; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation; D012639:Seeds / Q000737:chemistry; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D014197:Trees / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D001427:Bacterial Toxins; D003603:Cytotoxins; D010940:Plant Proteins; D052899:Pore Forming Cytotoxic Proteins; D011498:Protein Precursors; C073270:enterolobin protein, Enterolobium contortisiliquum; C032676:aerolysin