Development of a protein-chip containing Mitf for depigmenting agent screening.

초록

For the high-through-put-screening (HTS) of depigmenting agents, a protein chip was developed. Mitf (Microphthalmia-associated Transcription Factor), a key regulator in the melanogenesis, was produced as a fusion protein (Maltose Binding Protein-Mitf) in E. coli and was immobilized on the cyclodextrin-coated glass plate. Binding of oligo-DNA from E box of the promoter was monitored by fluorescence. The sequence of oligonucleotide-inhibitor affected the binding of the E-box fragment to Mitf, depending on the location of the sequence variation in the Ebox fragment. Changes in the CATGTG sequence reduced the binding of the E-box to Mitf, whereas no significant effect was observed when the sequence outside the CATGTG was varied. This result indicated that CATGTG is crucial for the binding of Mitf to E-box which initiates the transcription of pigmenting genes.