Hsp70(241–258), an octadecapeptide derived from the heat shock protein 70 (Hsp70) of rice (Oryza sativa
L. japonica), is a novel cationic -helical antimicrobial peptide (AMP) that contains four lysine, two arginine,
and two histidine residues. The antimicrobial activity of Hsp70(241–258) against Porphyromonas
gingivalis, a periodontal pathogen, and Candida albicans, an opportunistic fungal pathogen, was quantitatively
evaluated using a chemiluminescence method that measures ATP derived from viable cells. The
50% growth-inhibitory concentrations of Hsp70(241–258) against P. gingivalis and C. albicans cells were
63 M and 70 M, respectively. Hsp70(241–258) had little or no hemolytic activity even at 1 mM, and
showed negligible cytotoxicity up to 300 M. The degrees of calcein leakage from large unilamellar vesicles,
which mimic the membranes of Gram-negative bacteria, and 3,3-dipropylthiadicarbocyanine iodide
release from P. gingivalis cells induced by the addition of Hsp70(241–258) increased in a concentrationdependent
manner. When Hsp70(241–258) was added to calcein-acetoxymethyl ester-loaded C. albicans
cells, calcein release from the cells increased in a concentration-dependent manner. Flow cytometric
analysis also showed that the percentages of C. albicans cells stained with propidium iodide, a DNAintercalating
dye, increased as the concentration of Hsp70(241–258) added was increased. Therefore,
Hsp70(241–258) appears to exhibit antimicrobial activity against P. gingivalis and C. albicans through
membrane disruption. These results suggest that Hsp70(241–258) could be useful as a safe and potent
AMP against P. gingivalis and C. albicans in many fields of health care, especially in the control of oral
infections.
Hsp70(241–258), an octadecapeptide derived from the heat shock protein 70 (Hsp70) of rice (Oryza sativaL. japonica), is a novel cationic -helical antimicrobial peptide (AMP) that contains four lysine, two arginine,and two histidine residues. The antimicrobial activity of Hsp70(241–258) against Porphyromonasgingivalis, a periodontal pathogen, and Candida albicans, an opportunistic fungal pathogen, was quantitativelyevaluated using a chemiluminescence method that measures ATP derived from viable cells. The50% growth-inhibitory concentrations of Hsp70(241–258) against P. gingivalis and C. albicans cells were63 M and 70 M, respectively. Hsp70(241–258) had little or no hemolytic activity even at 1 mM, andshowed negligible cytotoxicity up to 300 M. The degrees of calcein leakage from large unilamellar vesicles,which mimic the membranes of Gram-negative bacteria, and 3,3-dipropylthiadicarbocyanine iodiderelease from P. gingivalis cells induced by the addition of Hsp70(241–258) increased in a concentrationdependentmanner. When Hsp70(241–258) was added to calcein-acetoxymethyl ester-loaded C. albicanscells, calcein release from the cells increased in a concentration-dependent manner. Flow cytometricanalysis also showed that the percentages of C. albicans cells stained with propidium iodide, a DNAintercalatingdye, increased as the concentration of Hsp70(241–258) added was increased. Therefore,Hsp70(241–258) appears to exhibit antimicrobial activity against P. gingivalis and C. albicans throughmembrane disruption. These results suggest that Hsp70(241–258) could be useful as a safe and potentAMP against P. gingivalis and C. albicans in many fields of health care, especially in the control of oralinfections.
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