본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Journal of microbiology and biotechnology v.15 no.2, 2005년, pp.254 - 258   피인용횟수: 7

In Vitro Formation of Protein Nanoparticle Using Recombinant Human Ferritin H and L Chains Produced from E. coli

RO HYEON SU    (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology   ); PARK HYUN KYU    (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology   ); KIM MIN GON    (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology   ); CHUNG BONG HYUN    (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology  );
  • 초록

    We have conducted in vitro reconstitution study of ferritin from its subunits FerH and FerL. For the reconstitution, FerH was produced from an expression vector construct in Escherichia coli and was purified from a heat treated cell extract by using one-step column chromatography. FerL was expressed as inclusion bodies. The denatured form of FerL was obtained by a simple washing step of the inclusion bodies with 3 M urea. The reconstitution experiment was conducted with various molar ratios of urea-denatured FerH and FerL to make the ferritin nanoparticle with a controlled composition of FerH and FerL. SDS-PAGE analysis of the reconstituted ferritins revealed that the reconstitution required the presence of more than 40 molar $\%$ of FerH in the reconstitution mixture. The assembly of the subunits into the ferritin nanoparticle was confmned by the presence of spherical particles with diameter of 10 nm by the atomic force microscopic image. Further analysis of the particles by using a transmission electron microscope revealed that the reconstituted particles exhibited different percentages of population with dense iron core. The reconstituted ferritin nanoparticles made with molar ratios of [FerH]/[FerL]=l00/0 and 60/40 showed that 80 to $90\%$ of the particles were apoferritin, devoid of iron core. On the contrary, all the particles formed with [FerH]/[FerL]=85/ 15 were found to contain the iron core. This suggests that although FerH can uptake iron, a minor portion of FerL, not exceeding $40\%$ at most, is required to deposit iron inside the particle.


  • 참고문헌 (19)

    1. Grace, J. E., M. E. Van Eden, and S. D. Aust. 2000. Production of recombinant human apoferritin heteromers. Arch. Biochem. Biophys. 384: 116- 122 
    2. Hempstead, P. D., S. J. Yewdall, A. R. Fernie, D. M. Lawson, P. J. Artymiuk, D. W. Rice, G. C. Ford, and P. M. Harrison. 1997. Comparison of the three-dimensional structures of recombinant human H and horse L ferritins at high resolution. J. Mol. Bol. 268: 424- 448 
    3. Hudson, A. J., S. C. Andrews, C. Hawkins. J. M. Williams, M. Izuhara, F. C. Meldrum, S. Mann, P. M. Harrison. and J. R. Guest. 1993. Overproduction, purification and characterization of the Escherichia coli ferritin. Eur. J. Biochem. 218: 985- 995 
    4. Kim, S. W., Y. H. Kim, and J. W. Lee. 2001. Thermal stability of human ferritin: Concentration dependence and enhanced stability of an N-terminal fusion mutant. Biochem. Biophys. Res. Commun. 289: 125-129 
    5. Meldrum, F. C, B. R. Heywood, and S. Mann. 1992. Magnetoferritin: In vitro synthesis of a novel magnetic protein. Science 257: 522- 523 
    6. Santambrogio, P., S. Levi, A. Cozzi, E. Rovida, A. Albertin, and P. Arosio. 1993. Production and characterization of recombinant heteropolymers of human ferritin H and L chains. J. Biol. Chem. 268: 12744- 12748 
    7. Von Darl, M., P. M. Harrison, and W. Bottke. 1994. Expression in Escherichia coli of a secreted invertebrate ferritin. Eur. J. Biochem. 222: 367- 376 
    8. Levi, S., A. Luzzago, G. Cesareni, A. Cozzi, F. Franceschinelli, A. Albertini, and P. Arosio. 1988. Mechanism of ferritin iron uptake: Activity of the H-chain and deletion mapping of the ferro-oxidase site. J. Biol. Chem. 263: 18086- 18092 
    9. Oh, K. S., Y. S. Park, and H. J. Sung. 2002. Expression and purification of an ACE-inhibitory peptide multimer from synthetic DNA in Escherichia coli. J. Microbiol. Biotechnol. 12: 59- 64 
    10. Lee, J., H. Y. Seo, E. S. Jeon, O. S. Park, K. M. Lee, C. U. Park, and K. S. Kim. 2001. Cooperative activity of subunits of human ferritin heteropolymers in Escherichia coli. J. Biochem. Mol. Biol. 34: 365- 370 
    11. Wong, K. K. W., T. Douglas, S. Gider, D. D. Awschalom, and S. Mann. 1998. Biomimetic synthesis and characterization of magnetic proteins magnetoferritin. Chem. Mater. 10: 279- 285 
    12. Wong, K. K. W., H. Colfen, N. T. Whilton, T. Dougals, and S. Mann. 1999. Synthesis and characterization of hydrophobic ferritin proteins. J. Inorg, Biochem. 76: 187- 195 
    13. Grady. J. K., Z. Zang, T. M. Laue, P. Arosio, and N. D. Chasteen. 2002. Characterization of the H- and L-subunit ratios of ferritins by sodium dodecyl sulfate-capillary gel electrophoresis. Anal. Biochem. 302: 263- 268 
    14. Oh, B. K., Y. K. Kim, Y. M. Bae, W. H. Lee, and J. W. Choi. 2002. Detection of Escherichia coli O157:H7 using imrnunosensor based on surface plasmon resonance. J. Microbiol. Biotechnol. 12: 780- 786 
    15. Levi, S., J. Salfeld, F. Franceschinelli, A. Cozzi, M. H. Dorner, and P. Arosio. 1989. Expression and structural and functional properties of human ferritin L-chain from Escherichia coli. Biochemistry 28: 5179- 5184 
    16. Levi, S., S. J. Yewdall, P. M. Harrison, P. Santambrogio, A. Cozzi, E. Rovida, A. Albertini, and P Arosio, 1992. Evidence that H- and L-chains have co-operative roles in the iron-uptake mechanism of human ferritin. Biochem. J. 288: 591- 596 
    17. Feng, L. and S. Damodaran. 2000. Two dimensional array of magnetoferritin on quarts surface. Thin Solid Films 365: 99-103 
    18. Harrison, P. M. and P. Arosio. 1996. The ferritins: Molecular properties, iron storage function and cellular regulation. Biochim. Biophys. Acta 1275: 161-203 
    19. Rucker, P., F. M. Torti, and S. V. Torti. 1997. Recombinant ferritin: Modulation of subunit stoichiometry in bacterial expression systems. Protein Eng. 10: 967- 973 
  • 이 논문을 인용한 문헌 (7)

    1. 2007. "" Journal of microbiology and biotechnology, 17(12): 2027~2032     
    2. 2007. "" Journal of microbiology and biotechnology, 17(10): 1695~1699     
    3. 2007. "" Journal of microbiology and biotechnology, 17(10): 1721~1726     
    4. 2007. "" Journal of microbiology and biotechnology, 17(4): 579~585     
    5. 2007. "" Journal of microbiology and biotechnology, 17(4): 638~643     
    6. 2008. "" Journal of microbiology and biotechnology, 18(2): 299~307     
    7. 2009. "" Journal of microbiology and biotechnology, 19(9): 911~917     

 저자의 다른 논문

  • CHUNG, BONG HYUN (1)

    1. 2005 "Production of Active Carboxypeptidase Y of Saccharomyces cerevisiae Secreted from Methylotrophic Yeast Pichia pastoris" Journal of microbiology and biotechnology 15 (1): 202~205    

 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