- 첨부된 파일이 없습니다.
- 조회 236
- 작성자 이태화
Cisperone-Mediated Protein Folding in vivo : A New Platform Technology for High-Throughput Protein Expression
- Date/Time : Mon August 25., 2003
- Speaker : 성 백 린
- Department of Biotechnology, Yonsei University, Seoul, KOREA
- Place : Life Science Bldg. #104
- For inquires : Professor Young-Chul Sung Dept. of Life Science
생명과학과 성영철 교수 (☎279-2294)
- Abstract -
A routine supply of functional proteinsemerges as a new bottleneck for various disciplines of post-genome research initiatives - high-throughput protein crystallization for structure determination and functional analysis, development of new drug screening methods in combination with in silico design, and development of various therapeutic proteins. A vexing problem for expression of recombinant protein in E. colihost is the inclusion body formation, precipitate of insoluble and inactive form of protein. Here, we report a new avenue for expression of proteins as soluble and functional forms by expeditingprotein folding in vivo.The ProFuse vector system capitalizes on a new chaperone molecule, which facilitates folding of target protein, when expressed as fusion to the chaperone. The chaperone functioning in cis, so christened with 'cisperone', is expected to work in a unique mechanism, distinct from the classical chaperones that work in trans. A varietyof proteins were tested and the ProFuse vector displayed extreme ability for increasing the solubility of the expressed proteins, far exceeding conventional fusion vectors based on fusion to thioredoxin or MBP. The protein could be used for new assay development for enzyme inhibitor screening. Alternatively, after cleavage with site-specific proteases, the target protein could be purified as native form, suitable for HT crystallization studies and therapeutic applications.