The pore-forming peptide of Entamoeba histolytica, the protozoan parasite causing human amoebiasis

Abstract

Amoebapore, the pore-forming peptide of E. histolytica has been isolated and its structure elucidated on the cDNA and protein level. The peptide is composed of 77 amino acid residues including six cysteine residues and has a molecular mass of 8244 Da. The primary translation product contains a signal sequence of 21 mostly hydrophobic amino acid residues. The active peptide has been located in the cytoplasmic granules of the amoebae. Circular dichroism spectroscopy revealed an all alpha-helical conformation and computer-aided secondary structure prediction yielded a structure of four helices. The helical conformation and three intramolecular disulfide bonds impart a highly compact and rigid structure upon the molecule. The activity of amoebapore, measured by a liposome depolarization assay, is resistant to heating at 100 degrees C in the absence of reducing agents. Synthetic peptides corresponding to the helices 1 and 3 exhibited pore-forming activity. Two minor, biologically active isoforms of amoebapore have amino acid sequence identity of 57% and 47%, respectively. Whereas amoebapore is a constituent of pathogenic E. histolytica isolates, nonpathogenic E. histolytica produce a structurally very similar peptide, the specific activity of which is approximately one third that of amoebapore. The biological significance of amoebapore for the pathogenicity of E. histolytica and specifically for its cytolytic activity remains to be determined.