Application of hybrid helix method in structural biology = 하이브리드 헬릭스 방법을 응용한 단백질 구조 연구

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Artificial protein assemblies require a method to conjugate two or more proteins in a stable and predictable structure. “Shared helix method” is useful for protein engineering that fuses the exposed alpha helices from two different protein components rigidly and predictably. In this study, the successful design of these novel fusion proteins, Putrescine aminotransferase (YgjG) complexes, was confirmed by X-ray crystallography. Also, we applied shared helix method to structural analysis of small target proteins (~<100kDa) using cryo-EM. To overcome size limitations in cryo-EM, we selected glutamate dehydrogenase (GDH) as a scaffold protein to determine the structure of small target proteins and the protein assemblies were successfully constructed using shared helix method. This application does not need to optimize the extra sequence of linker to connect each protein and forms stable complex conformation. Since, using the high molecule protein and small target protein complex, which was fused by alpha helix fusion method, is capable to assemble the cryo-EM and may improve to be convenient tool for cryo-EM analysis of small target protein.
Advisors
Lee, Jie-Ohresearcher이지오researcher
Description
한국과학기술원 :화학과,
Publisher
한국과학기술원
Issue Date
2018
Identifier
325007
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 화학과, 2018.8,[v, 72 p. :]

Keywords

Alpha helix▼afusion method▼arepeat protein▼aprotein complex▼acrystallography▼acryo-EM; 알파 헬릭스▼a융합 방법▼a반복 단백질▼a단백질 복합체▼a단백질 결정학▼a저온전자현미경

URI
http://hdl.handle.net/10203/265487
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=867928&flag=dissertation
Appears in Collection
CH-Theses_Ph.D.(박사논문)
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