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In previous study, it was reported that the 30K protein originating from silkworm, Bombyx mori, inhibited apoptosis in mammalian cells. In this works, we observed a novel property of 30K protein; cell-penetrating property. When recombinant 30Kc19 produced from E.coli was added to
culture medium, it was shown that 30K protein was internalized into the living human cell lines. The degree of internalization was increased in a dose-dependent manner and this process was occurred within short period of time (< 1 hr). Moreover, it was examined that dextran sulfate, a
kind of poly-anionic material, in the culture medium inhibited the internalization of 30K protein. To examine a cellular localization of 30K protein, immunofluorescence (IF) assay was performed using confocal microscopy. It was demonstrated that 30K protein was bound onto the cell membrane within 10 min and internalized gradually. According to these IF results, the internalization was completed within about 1 hr. Consequently, we conclude that 30K protein is a novel cell-penetrating proteins such as TAT or Antp because these properties are similar with
that of the other cell-penetrating proteins. Cell-penetrating protein and its protein transduction domain (PTD) have been used to deliver macromolecules and even small molecules for their functions inside the cells. Therefore, our results support that 30K protein is anticipated to be
used as new tool to deliver macromolecules into the living cells.