Abstract
Nisin is a food preservative produced by Lactococcus lactis subsp. lactis. Previous blood biochemical research revealed that nisin has physiological effects in mammals; although the site of action has yet to be identified, keratinocytes have been proposed as a possible target. In this study, we investigated whether nisin affects keratinocytes by examining the effects on eukaryotic intermediate filaments in HaCaT human keratinocytes. Treatment with 93 μg/ml nisin for 24 h decreased the localization of the intermediate filament proteins cytokeratin (CK)5 and CK17 at the cell periphery, which were distributed in a limited area in a ring- or net-like shape. However, this was not observed upon treatment for 6 h. The results of a serial dilution assay revealed that the effect on CK17 localization depends on the nisin concentration and were observed at ≥47 μg/ml. Moreover, this effect was partially blocked by treatment with the calcium channel blocker bepridil. Thus, despite the long history of nisin as being safe for humans, it has measurable effects on the keratinocyte cytoskeleton. Our findings also indicate that CK5 and CK17 can serve as markers for evaluating the effects of nisin on keratinocytes.
Highlights
Bacteriocins are antimicrobial peptides produced by various bacteria
We evaluated the effects of nisin on keratinocytes by examining the changes in the distribution of eukaryotic intermediate filament proteins, namely CK5 and CK17, in HaCaT cells, which is a widely used immortal human keratinocyte cell line that has uniform epithelial architecture when transplanted into nude mice (Boukamp et al 1988)
We demonstrated that nisin treatment altered the intermediate filament distribution in keratinocytes
Summary
Bacteriocins are antimicrobial peptides produced by various bacteria. Nisin is a type of bacteriocin produced by some strains of Lactococcus lactis subsp. lactis. We speculated that nisin acts on keratinocytes prior to its digestion by proteases. To test this hypothesis, we evaluated the effects of nisin on keratinocytes by examining the changes in the distribution of eukaryotic intermediate filament proteins, namely CK5 and CK17, in HaCaT cells, which is a widely used immortal human keratinocyte cell line that has uniform epithelial architecture when transplanted into nude mice (Boukamp et al 1988). Changes in CK distribution are correlated with cell growth and epithelialization, cell migration, and cytoarchitecture (Paladini et al 1996; Foisner 1997) Due to their ubiquitous distribution in the cytoplasm, intermediate filament proteins can be used for detection of previously undetected alterations brought about by nisin
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