대한화장품학회지 (Journal of the Society of Cosmetic Scientists of Korea)

  • 제30권4호
  • /
  • Pages.479-483
  • /
  • 2004
  • /
  • 1226-2587(pISSN)
  • /
  • 2288-9507(eISSN)
대한화장품학회 (Society of Cosmetic Scientists of Korea)
권호 표지

전사인자 저해제 통한 미백제 탐색용 단백질 칩 제작

Manufacturing Protein-DNA Chip for Depigmenting Agent Screening

  • 한정선 (인하대학교 공과대학 생명화학공학부 생물공학과) ;
  • 곽은영 (인하대학교 공과대학 생명화학공학부 생물공학과) ;
  • 이향복 (인하대학교 공과대학 생명화학공학부 생물공학과) ;
  • 신정현 (인하대학교 의과대학 피부과) ;
  • 백승학 (한국생명공학연구원 바이오나노센터) ;
  • 정봉현 (한국생명공학연구원 바이오나노센터) ;
  • 김은기 (인하대학교 공과대학 생명화학공학부 생물공학과)
  • Han Jung-Sun (Department of Biological Engineering, Inha University) ;
  • Kwak Eun-Young (Department of Biological Engineering, Inha University) ;
  • Lee Hyang-Bok (Department of Biological Engineering, Inha University) ;
  • Shin Jlung-Hyun (Department of Dermatology, College of Medicine, Inha University) ;
  • Baek Seung-Hak (Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Chung Bong-Hyun (Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim Eun-Ki (Department of Biological Engineering, Inha University)
  • 발행 : 2004.12.01
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초록

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.

키워드

참고문헌

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  2. H. Aoki and O. Moro, Involvement of microphthal-mia-associated transcription factor (MITF) in expression of human melanocortin-1 receptor (MC1R), Life Sci., 71, 2171 (2002)
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  7. 특허: MBP chip을 이용한 MITF 전사인자 저해제의 고속탐색방법 개발(특허출원중)