Science in China Series B: Chemistry

, Volume 52, Issue 9, pp 1388–1393 | Cite as

Separation/enrichment of the low-content high molecular weight natural protein using protein-imprinted polymers with ARPCs

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Abstract

We introduce a new method for separation/enrichment of the low-content cellular protein in high molecular weight on the basis of molecular imprinting. The template protein, bacterial cloned immunoglobulin binding protein (BiP), was selectively assembled with assistant recognition polymer chains (ARPCs) from their library, which consists of numerous limited length polymer chains with randomly distributed recognition and immobilizing sites. The assemblies of proteins and ARPCs were adsorbed by porous polymeric beads and immobilized by cross-linking polymerization. After the template was removed, the synthesized imprinted polymer was used to adsorb authentic BiP from endoplasmic reticulum (ER) extract, and its proportional content was enriched 45 times. It is the first time that the low-content cellular natural protein, whose molecular weight reaches 78 kDa, is enriched by molecular imprinting.

Keywords

molecular imprinting technique assistant recognition polymer chains (ARPCs) bacterial cloned protein high molecular weight protein 
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References

  1. 1.
    Scopes R K. Protein Purification: Principles and Practice. Berlin: Springer-Verlag, 1982. 373–379Google Scholar
  2. 2.
    Deutscher M P. Guide to Protein Purification. San Diego: Academic Press, 1990. 339–343Google Scholar
  3. 3.
    Anderson L, Sellergren B, Mosbach K. Imprinting of amino acid derivatives in macroporous polymers. Tetrahedron Lett, 1984, 25: 5211–5214CrossRefGoogle Scholar
  4. 4.
    Vlatakis G, Andersson L, Mueller R, Mosbach K. Drug assay using antibody mimics made by molecular imprinting. Nature, 1993, 361: 645–647CrossRefGoogle Scholar
  5. 5.
    Wulff G. Molecular imprinting in cross-linked materials with the aid of molecular templates—A way towards artificial antibodies. Angew Chem Int Ed Engl, 1995, 34: 1812–1932CrossRefGoogle Scholar
  6. 6.
    Wulff G. Enzyme-like catalysis by molecularly imprinted polymers. Chem Rev, 2002, 102: 1–27CrossRefGoogle Scholar
  7. 7.
    Guo T Y, Xia Y Q, Hao G J, Song M D, Zhang B H. Adsorptive separation of hemoglobin by molecularly imprinted chitosan beads. Biomaterials, 2004, 25: 5905–5912CrossRefGoogle Scholar
  8. 8.
    Burow M, Minoura N. Molecular imprinting: synthesis of polymer particles with antibody-like binding characteristics for glucose oxidase. Biochem Biophys Res Commun, 1996, 227: 419–422CrossRefGoogle Scholar
  9. 9.
    Hirayama K, Burow M, Morikawa, Minoura N. Synthesis of polymer-coated silica particles with specific recognition sites for glucose oxidase by the molecular imprinting technique. Chem Lett, 1998, 8: 731–732CrossRefGoogle Scholar
  10. 10.
    Haupt K. Creating a good impression. Nat Biotechnol, 2002, 20: 884–885CrossRefGoogle Scholar
  11. 11.
    Guo M J, Zhao Z, Fan Y G, Wang C H, Shi L Q, Xia J J, Long Y, Mi H F. Protein-imprinted polymer with immobilized assistant recognition polymer chains. Biomaterials, 2006, 27: 4381–4387CrossRefGoogle Scholar
  12. 12.
    Long Y, Xing X C, Han R F, Sun Y, Wang Y, Zhao Z, Mi H F. Two-step purification of low-content cellular protein using protein-imprinted polymers. Anal Biochem, 2008, 380: 268–275CrossRefGoogle Scholar
  13. 13.
    Han R F, Xing X C, Wang Y, Long Y, Sun Y, Zhao Z, Mi H F. Separation/enrichment of active natural low content protein using protein imprinted polymer. J Chromatogr B, 2008, 873:113–118CrossRefGoogle Scholar
  14. 14.
    Zhao Z, Wang C H, Guo M J, Shi L Q, Fan Y G, Long Y, Mi H F. Molecular imprinted polymer with cloned bacterial protein template enriches authentic target in cell extract. FEBS Lett, 2006, 580: 2750–2754CrossRefGoogle Scholar
  15. 15.
    Haas I G, Wabl M. Immunoglobulin heavy chain binding protein. Nature, 1983, 306: 387–389CrossRefGoogle Scholar
  16. 16.
    Munro S, Pelham H R. An Hsp70-like protein in the ER: identity with the 78 kD glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell, 1986, 46: 291–300CrossRefGoogle Scholar
  17. 17.
    Haas I G, Meo T. cDNA cloning of the immunoglobulin heavy chain binding protein. Proc Natl Acad Sci USA, 1988, 85: 2250–2254CrossRefGoogle Scholar
  18. 18.
    Zhu X, Zhao X, Burkholder W F, Gragerov A, Ogata CM, Gottesman M E, Hendrickson W A. Structural analysis of substrate binding by the molecular chaperone DnaK. Science, 1996, 272: 1606–1614CrossRefGoogle Scholar
  19. 19.
    Rudiger S, Buchberger A, Bukau B. Interaction of Hsp70 chaperones with substrates. Nat Struct Biol, 1997, 4: 342–349CrossRefGoogle Scholar
  20. 20.
    Rudiger S, Mayer M P, Schneider-Mergener J, Bukau B. Modulation of substrate specificity of the DnaK chaperone by alteration of a hydrophobic arch. J Mol Biol, 2000, 304: 245–251CrossRefGoogle Scholar
  21. 21.
    Kassenbrock C K, Kelly R B. Interaction of heavy chain binding protein (BiP/GRP78) with adenine nucleotides. EMBO J, 1989, 8: 1461–1467Google Scholar
  22. 22.
    Zhang X B, Li H, Zhang W Q, Wang Y, Wu D, Mi H F. The mouse FKBP23 binds to BiP in ER and the binding of C-terminal domain is interrelated with Ca2+ concentration. FEBS lett, 2004, 559(1–3): 57–60CrossRefGoogle Scholar
  23. 23.
    Borgeson C E, Bowman B J. Isolation and characterization of the Neurospora crassa endoplasmic reticulum. J Bacteriol, 1983, 156: 362–368Google Scholar
  24. 24.
    Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem, 1976, 72: 248–254CrossRefGoogle Scholar

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© Science in China Press and Springer Berlin Heidelberg 2009

Authors and Affiliations

  1. 1.Biochemical Section of Key Laboratory of Functional Polymer Materials, Ministry of Education of China, Institute of Polymer ChemistryNankai UniversityTianjinChina

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