본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Bacterial ${\beta}$-Lactamase Fragment Complementation Strategy Can Be Used as a Method for Identifying Interacting Protein Pairs

Park, Jong-Hwa   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  ); Back, Jung-Ho   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  ); Hahm, Soo-Hyun   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  ); Shim, Hye-Young   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  ); Park, Min-Ju   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  ); Ko, Sung-Il   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  ); Han, Ye-Sun   (Department of Advanced Technology Fusion and Bio-Molecular Informatics Center, Konkuk UniversityUU0000050  );
  • 초록

    We investigated the applicability of the TEM-l ${\beta}$ -lactamase fragment complementation (BFC) system to develop a strategy for the screening of protein-protein interactions in bacteria. A BFC system containing a human Fas-associated death domain (hFADD) and human Fas death domain (hFasDD) was generated. The hFADD-hFasDD interaction was verified by cell survivability in ampicillin-containing medium and the colorimetric change of nitrocefin. It was also confirmed by His pull-down assay using cell lysates obtained in selection steps. A coiled-coil helix coiled-coil domain-containing protein 5 (CHCH5) was identified as an interacting protein of human uracil DNA glycosylase (hUNG) from the bacterial BFC cDNA library strategy. The interaction between hUNG and CHCH5 was further confirmed with immunoprecipitation using a mammalian expression system. CHCH5 enhanced the DNA glycosylase activity of hUNG to remove uracil from DNA duplexes containing a U/G mismatch pair. These results suggest that the bacterial BFC cDNA library strategy can be effectively used to identify interacting protein pairs.


  • 주제어

    ${\beta}$-Lactamase fragment complementation .   hFADD .   hFasDD .   nitrocefin .   cDNA library .   human uracil DNA glycosylase .   coiled-coil helix coiled-coil domain-containing protein 5.  

  • 참고문헌 (29)

    1. Drewes, G. and T. Bouwmeester. 2003. Global approaches to protein-protein interactions. Curr. Opin. Cell Biol. 15: 199-205 
    2. Ghosh, I., A. D. Hamilton, and L. Regan. 2000. Antiparallel leucine zipper-directed protein reassembly: Application to the green fluorescent protein. J. Am. Chem. Soc. 122: 5658-5659 
    3. Hill, J. M., G. Morisawa, T. Kim, T. Huang, Y. Wei, Y. Wei, and M. H. Werner. 2003. Identification of an expanded binding surface on the FADD death domain responsible for interaction with CD95/Fas. J. Biol. Chem. 279: 1474-1481 
    4. Nilsen, H. and H. E. Krokan. 2001. Base excision repair in a network of defense and tolerance. Carcinogenesis 22: 987-998 
    5. Paulmurugan, R. and S. S. Gambhir. 2003. Monitoring protein-protein interactions using split synthetic Renilla luciferase protein-fragment-assisted complementation. Anal. Chem. 75: 1584-1589 
    6. Rossi, F. M., B. T. Blakely, and H. M. Blau. 2000. Interaction blues: Protein interactions monitored in live mammalian cells by beta-galactosidase complementation. Trends Cell Biol. 10: 119-122 
    7. Wehrman, T., B. Kleaveland, J. H. Her, R. F. Balint, and H. M. Blau. 2002. Protein-protein interactions monitored in mammalian cells via complementation of beta-lactamase enzyme fragments. Proc. Natl. Acad. Sci. USA 99: 3469-3474 
    8. Michnick, S. W. 2003. Protein fragment complementation strategies for biochemical network mapping. Curr. Opin. Biotech. 14: 610-617 
    9. Campbell, R. E. 2004. Realization of beta-lactamase as a versatile fluorogenic reporter. Trends Biotechnol. 22: 208-211 
    10. Ito, T., K. Tashiro, S. Muta, R. Osawa, T. Chiba, M. Nishizawa, K. Yamamoto, S. Kuhara, and Y. Sakaki. 2000. Toward a protein-protein interaction map of the budding yeast: A comprehensive system to examine two-hybrid interactions in all possible combinations between the yeast proteins. Proc. Natl. Acad. Sci. USA 97: 1143-1147 
    11. Jim, Y. T., T. U. Kim, and H. S. Baik. 2006. Characterization of extended spectrum $\beta$-lactamase genotype TEM, SHV, and CTX-M producing Klebsiella pneumoniae isolated from clinical specimens in Korea. J. Microbiol. Biotechnol. 16: 889-895     
    12. Kavil, B., O. Sundheim, M. Kbari, M. Otterlei, H. Nilsen, F. Skorpen, P. A. Aas, L. Hagen, H. E. Krokan, and G. Slupphaug. 2002. hUNG2 is the major repair enzyme for removal of uracil from U:A matches, U:G mismatches and U in single stranded DNA, with hSMUNG1 as a broad specificity backup. J. Biol. Chem. 277: 39926-39936 
    13. Nord, O., A. Gustrin, and P. Nygren. 2005. Fluorescent detection of $\beta$-lactamase activity in living Escherichia coli cells via esterase supplementation. FEMS Microbiol. Lett. 242: 73-79 
    14. Otterlei, M., E. Warbrick, T. A. Nagelhus, T. Haug, G. Slupphaug, M. Akbari, P. A. Aas, K. Steinsbekk, O. Bakke, and H. E. Krokan. 1999. Post-replicative base excision repair in replication foci. EMBO J. 18: 3834-3844 
    15. Pelletier, J. N., K. M. Arndt, A. Pluckthun, and S. W. Michnick. 1999. An in vivo library-versus-library selection of optimized protein-protein interactions. Nat. Biotechnol. 17: 683-690 
    16. Fields, S. and O. Song. 1989. A novel genetic system to detect protein-protein interactions. Nature 340: 245-246 
    17. Kang, J. W. and Y. J. Kim. 2005. Enzymatic and energetic properties of an aerobic respiratory chain-linked NADH oxidase system in marine bacterium Vibrio natriegens. J. Microbiol. Biotechnol. 15: 1080-1086     
    18. Weldhangen, G. F., B. H. Kim, C. H. Cho, and S. H. Lee. 2006. Definitive nomenclature of GES/IBC-type extendedspectrum $\beta$-lactamases. J. Microbiol. Biotechnol. 16: 1837-1840     
    19. Spotts, J. M., R. E. Dolmetsch, and M. E. Greenberg. 2002. Time-lapse imaging of a dynamic phosphorylationdependent protein-protein interaction in mammalian cells. Proc. Natl. Acad. Sci. USA 99: 15142-15147 
    20. Krokan, H. E., F. Drablos, and G. Slupphaug. 2002. Uracil in DNA-occurrence, consequences and repair. Oncogene 21: 8935-8948 
    21. Pollok, B. A. and R. Heim. 1999. Using GFP in FRET-based applications. Trends Cell Biol. 9: 57-60 
    22. Moore, J. T., S. T. Davis, and I. K. Dev. 1997. The development of beta-lactamase as a highly versatile genetic reporter for eukaryotic cells. Anal. Biochem. 247: 203-209 
    23. Nobrega, M. P., S. C. Bandeira, J. Beers, and A. Tzagoloff. 2002. Characterization of COX19, a widely distributed gene required for expression of mitochondrial cytochrome oxidase. J. Biol. Chem. 277: 40206-40211 
    24. Galaneau, A., M. Primeau, L. E. Trudeau, and S. W. Michnick. 2002. Beta-lactamase protein fragment complementation assays as in vivo and in vitro sensors of protein interactions. Nat. Biotechnol. 20: 619-622 
    25. Triepels, R., L. van den Heuvel, J. Loeffen, R. Smeets, F. Trijbels, and J. Smeritink. 1998. The nuclear-encoded human NADH: Ubiquinone oxidoreductase NDUFA8 subunit: cDNA cloning, chromosomal localization, tissue distribution, and mutation detection in complex-I-deficient patients. Hum. Genet. 103: 557-563 
    26. Jin, C., A. M. Myers, and A. Tzagoloff. 1997. Cloning and characterization of MRP10, a yeast gene coding for a mitochondrial ribosomal protein. Curr. Genet. 31: 228-234 
    27. Remy, I. and S. W. Mitchnick. 2001. Visualization of biochemical networks in living cells. Proc. Natl. Acad. Sci. USA 98: 7678-7683 
    28. Zlokarnik, G., P. A. Negulescu, T. E. Knapp, L. Mere, N. Burres, L. Feng, M. Whitney, K. Roemer, and R. Y. Tsien. 1998. Quantization of transcription and clonal selection of single living cells with beta-lactamase as reporter. Science 279: 84-88 
    29. Krokan, H. E., M. Otterlei, H. Nilsen, B. Kavli, F. Skorpen, S. Anderson, C. Skjelbred, M. Akbari, P. A. Aas, and G. Slupphaug. 2001. Properties and functions of human uracil- DNA glycosylase from the UNG gene. Prog. Nucleic Acid Res. Mol. Biol. 68: 365-386 

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
유료다운로드

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기