Cell binding of pneumolysin to target cells is an important step in the lysis of cells by this toxin. We sought to locate the cell-binding region of pneumolysin. Deletion of the six C-terminal amino acids decreased cell-binding activity by 98%. Furthermore, mutagenesis of an amino acid near the C-terminus decreased the cell-binding activity of full-length pneumolysin by 90%. The C-terminus of pneumolysin has an important role in cell-binding activity.