Miri Seiberg

Summary: The protease‐activated receptor‐2 (PAR‐2) is a seven transmembrane G‐protein‐coupled receptor that could be activated by serine protease cleavage or by synthetic peptide agonists. We showed earlier that activation of PAR‐2 with Ser‐Leu‐Ile‐Gly‐Arg‐Leu‐NH₂(SLIGRL), a known PAR‐2 activating peptide, induces keratinocyte phagocytosis and increases skin pigmentation, indicating that PAR‐2 regulates pigmentation by controlling phagocytosis of melanosomes. Here, we show that Leu‐Ile‐Gly‐Arg‐NH₂(LIGR) can also induce skin pigmentation. Both SLIGRL and LIGR increased melanin deposition in vitro and in vivo, and visibly darkened human skins grafted onto severe combined immuno‐deficient (SCID) mice. Both SLIGRL and LIGR stimulated Rho‐GTP activation resulting in keratinocyte phagocytosis. Interestingly, LIGR activates only a subset of the PAR‐2 signaling pathways, and unlike SLIGRL, it does not induce inflammatory processes. LIGR did not affect many PAR‐2 signaling pathways, including [Ca²⁺] mobilization, cAMP induction, the induction of cyclooxgenase‐2 (COX‐2) expression and the secretion of prostaglandin E2, interleukin‐6 and ‐8. PAR‐2 siRNA inhibited LIGR‐induced phagocytosis, indicating that LIGR signals via PAR‐2. Our data suggest that LIGR is a more specific regulator of PAR‐2‐induced pigmentation relative to SLIGRL. Therefore, enhancing skin pigmentation by topical applications of LIGR may result in a desired tanned‐like skin color, without enhancing inflammatory processes, and without the need of UV exposure.

We have recently shown that soybean‐derived serine protease inhibitors and soybean extracts alter skin pigmentation, suggesting that soymilk could be used as a natural alternative to skin l...

The epidermal–melanin unit is composed of one melanocyte and approximately 36 neighboring keratinocytes, working in synchrony to produce and distribute melanin. Melanin is synthesized in melanosomes, transferred to the dendrite tips, and translocated into keratinocytes, forming caps over the keratinocyte nuclei. The molecular and cellular mechanisms involved in melanosome transfer and the keratinocyte–melanocyte interactions required for this process are not yet completely understood. Suggested mechanisms of melanosome transfer include melanosome release and endocytosis, direct inoculation (‘injection’), keratinocyte–melanocyte membrane fusion, and phagocytosis. Studies of the keratinocyte receptor protease‐activated receptor‐2 (PAR‐2) support the phagocytosis theory. PAR‐2 controls melanosome ingestion and phagocytosis by keratinocytes and exerts a regulatory role in skin pigmentation. Modulation of PAR‐2 activity can enhance or decrease melanosome transfer and affects pigmentation only when there is keratinocyte–melanocyte contact. Moreover, PAR‐2 is induced by UV irradiation and inhibition of PAR‐2 activation results in the prevention of UVB‐induced tanning. The role of PAR‐2 in mediating UV‐induced responses remains to be elucidated.

The protease-activated receptor-2 (PAR-2) belongs to the family of seven transmembrane domain receptors, which are activated by the specific enzymatic cleavage of their extracellular amino termini. Synthetic peptides corresponding to the tethered ligand domain (SLIGRL in mouse, SLIGKV in human) can activate PAR-2 without the need for receptor cleavage. PAR-2 activation is involved in cell growth, differentiation and inflammatory processes, and was shown to affect melanin and melanosome ingestion by human keratinocytes. Data presented here suggest that PAR-2 activation may regulate human keratinocyte phagocytosis. PAR-2 activation by trypsin, SLIGRL or SLIGKV increased the ability of keratinocytes to ingest fluorescently labeled microspheres or E. coli K-12 bioparticles. This PAR-2 mediated increase in keratinocyte phagocytic capability correlated with an increase in actin polymerization and α-actinin reorganization, cell surface morphological changes and increased soluble protease activity. Moreover, addition of serine protease inhibitors downmodulated both the constitutive and the PAR-2 mediated increases in phagocytosis, suggesting that serine proteases mediate this functional activity in keratinocytes. PAR-2 involvement in keratinocyte phagocytosis is a novel function for this receptor.