지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
초록· 키워드
오류제보하기
Parkin, known as an E3 ubiquitin ligase, has essential role in protein quality control, and its severe dysfunction leads to neurodegenerative disorders. Human Parkin was excessively degraded when expressed in Escherichia coli under the conventional induction condition (37℃ culture condition with 0.5 mM IPTG). To optimize the induction and culture conditions for recombinant human Parkin and develop a rapid method for the Parkin purification, we expressed Parkin by using pGEX system at the different culture temperatures and IPTG concentrations. The intact Parkin protein was purified to approximately 90% purity with suitable amounts of protein under the optimal culture condition (25℃ with 0.01 mM IPTG). Additionally, we constructed various parkin plasmids with different tagging systems and investigated their expression patterns in HEK293 cells. We found that the proteolytically sensitive site is localized within a ubiquitin-like domain of Parkin. This study developes a method for generating useful reagents to investigate biochemical properties of Parkin.
목차
Abstract
재료 및 방법
결과
고찰
요약
감사의 글
문헌
참고문헌 (26)
참고문헌 신청
Ardley H. C., C. C. Hung, and P. A. Robinson. 2005. The aggravating role of the ubiquitin-proteasome system in neurodegeneration. FEBS Lett. 579, 571-576.
Cyr D. M., J. Hohfeld, and C.Patterson. 2002. Protein quality control: U-box-containing E3 ubiquitin ligases join the fold. Trends Biochem Sci. 27, 368-375.
Tanaka K. and T. Chiba. 1998. The proteasome: a protein-destroying machine. Genes Cells. 3, 499-510.
Hershko A. and A. Ciechanover. 1998. The ubiquitin system. Annu Rev Biochem. 67, 425-479.
Frydman J. 2001. Folding of newly translated proteins in vivo: the role of molecular chaperones. Annu Rev Biochem. 70, 603-647.
Cyr D. M., J. Hohfeld, and C.Patterson. 2002. Protein quality control: U-box-containing E3 ubiquitin ligases join the fold. Trends Biochem Sci. 27, 368-375.
Tanaka K. and T. Chiba. 1998. The proteasome: a protein-destroying machine. Genes Cells. 3, 499-510.
Hershko A. and A. Ciechanover. 1998. The ubiquitin system. Annu Rev Biochem. 67, 425-479.
Frydman J. 2001. Folding of newly translated proteins in vivo: the role of molecular chaperones. Annu Rev Biochem. 70, 603-647.
함께 읽어보면 좋을 논문
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UCI(KEPA) : I410-ECN-0101-2009-470-015375916