MITF는 미백관련 유전자의 대표적인 조절 인자 단백질로서 미백관련 유전자의 E-box와의 결합정도를 단백질 칩을 이용하여 측정하였다. 융합 단백질 형태의 MITF를 유리 칩에 고정시켰고 E-box를 포함하는 DNA oligomer가 결합하는 것을 확인하였다. 형광법, SPR (surface plasmon resonance), SPRi (surface plasmon resonance imaging)방법 중 형광법이 가장 효과적이었으며, DNA 저해제를 사용시 결합이 감소하는 것을 확인하였다. 이 결과 MITF를 이용한 미백원료의 고속스크리닝(HTS)의 가능성을 보여주었다.
An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.
