표제지
논문개요
목차
Ⅰ. 서론 13
1. HTRA를 타겟으로 하는 형광 기질 합성 13
2. 신경세포주에서 분화상태에 따른 HTRA 활성변화의 탐색 15
3. HTRA를 타겟으로 하는 cyclic activity-based probe의 합성 17
Ⅱ. 결과 및 고찰 19
1. HTRA를 타겟으로 하는 형광 기질 합성 19
1) 테트라펩타이드 기반 라이브러리 합성 19
2) Enzymatic assay 20
2. 신경세포주에서 분화상태에 따른 HTRA 활성변화의 탐색 22
1) Cell differentiation 22
2) Selectivity in SH-SY5Y/SK-N-SH 23
3) Live cell imaging 24
3. HTRA를 타겟으로 하는 Cyclic Activity-based probe의 합성 26
1) hexa-peptide library의 합성 26
2) Valine-DPP-DCA 합성 26
3) Valine-DPP-DCA를 이용한 peptide cyclization 27
Ⅲ. 결론 30
Ⅳ. 실험 32
1. 실험기기 및 시약 32
1) 실험기기 32
2) 시약 32
2. 실험방법 33
1) HTRA를 타겟으로 하는 형광 기질 합성 33
2) 신경세포주에서 분화상태에 따른 HTRA 활성변화의 탐색 39
3) Cyclic Activity-based probe의 합성 42
참고문헌 47
Abstract 49
Table 1. Sequence of fluorogenic substrate for HTRA 19
Table 2. sequence of hexa-peptide library 26
Figure 1. Structure of 7-amino-4-methylcoumarin 14
Figure 2. Spectrum of 7-amino-4-methylcoumarin 14
Figure 3. SH-SY5Y cells undergo morphological changes during RA-induced differentiation (A) Undifferentiated SH-SY5Y cells cultured in complete growth... 15
Figure 4. Structure of Cyclosporine 17
Figure 5. Structural features of Ahp-cyclodepsipeptidesfor serine protease inhibition a) Chemical structures of the natural product Ahp-cyclodepsipeptides... 18
Figure 6. Measurement of signal change of fluorescent substrate at various HTRA2 concentrations 21
Figure 7. Cell differentiation of SH-SY5Y 22
Figure 8. Cell differentiation of SK-N-SH 22
Figure 9. Structure of HTRA probe (DS-01-94) 23
Figure 10. 2nd generation probe with SH-SY5Y/SK-N-SH[이미지참조] 24
Figure 11. Cell differentiation of SK-N-SH (control) 25
Figure 12. Cell differentiation of SK-N-SH (ATRA 최종농도 10µM) 25
Figure 13. structure of cyclic-activity based peptide probe 29
Scheme 1. synthesis of fluorogenic substrate for HTRA 19
Scheme 2. synthesis of cyclic-activity based peptide probe 28