융합 페리틴의 요소 농도에 따른 재접힘 특성에 관한 연구

Fusion ferritin$(F_H+F_L)$, an iron-binding protein, was purified from recombinant E. coli by two-step sonications with urea. Unfolded ferritin was refolded by gel filtration chromatography with various concentration of urea. 50 mM Tris-HCl(pH 8.0) buffers with 1 M to 4 M urea were used in GFC. Objective was to characterize the structure change with urea concentration. Molecular weight was determined using GF-HPLC and RP-HPLC was used to quantify the unfolded and refolded proteins.

세포 전처리 과정으로 융합 ferritin을 2단계 세포파쇄하여 주입시료로 이용하였으며, 변성된 단백질용액을 젤 여과 크로마토그래피를 통하여 urea의 농도를 희석시켜 재접힘 시켰다. 여기서 분취한 시료를 역상 크로마토그래피에 적용한 결과 FPLC 크로마토그램에서 뒤에 분취한 시료일수록 재접힘 정도가 높은 것으로 나타났다.