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Each of the four arginine residues in the HM-1 killer toxin was replaced by alanine using site-directed mutagenesis. The polymerase chain reaction (PCR)-constructed mutant gene was successfully expressed in HM-1 toxin resistant Saccharomyces cerevisiae. Among four HM-1 toxin analogues, R82A HM-1 toxin and R86A HM-1 toxin lost killer activity, while R61A HM-1 toxin and R85A HM-1 toxin retained activity. These results strongly indicate the importance of the arginine residues at positions 82 and 86 which are located in the C-terminal region of the HM-1 toxin for the action of killer activity.

The Involvement of Two Specific Arginine Residues in the Action of HM-1 Killer Toxin was Deduced from Site-Directed Mutagenesis.