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Evaluation of surface hydrophobicity of immobilized protein with a surface plasmon resonance sensor

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Abstract

Immobilization is widely used to isolate agglutinative and associative proteins with large hydrophobic surfaces. Surface hydrophobicities of immobilized proteins were quantified by measuring the adsorption amounts of Triton X-100 as a hydrophobic probe with a biosensor that utilizes the phenomena of surface plasmon resonance (SPR). We measured SPR signal changes derived from adsorption of Triton X-100 to five kinds proteins and calculated the monolayer adsorption capacity using the Brunauer–Emmett–Teller equation, partly modified with a term for correcting an influence of the net charge of immobilized protein. SPR signal changes obtained by this method correlated with the values of surface hydrophobicities obtained by conventional assay using a hydrophobic probe. Thus this measuring method using an SPR sensor and Triton X-100 is expected to be a tool for quantifying surface hydrophobicities of immobilized proteins.

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References

  • Alizadeh-Pasdar N, Li-Chan ECY (2000) Comparison of protein surface hydrophobicity measured at various pH values using three different fluorescent probes. J. Agric. Food Chem. 48: 328–334.

    PubMed  Google Scholar 

  • Bam NB, Cleland JL, Randolph TW (1996) Molten globular intermediate of recombinant human growth hormone: stabilization and surfactants. Biotechnol. Prog. 12: 801–809.

    PubMed  Google Scholar 

  • Clark EDB (2001) Protein refolding for industrial processes. Curr. Opin. Biotechnol. 12: 202–207.

    PubMed  Google Scholar 

  • Gu T, Rupprecht H, Galera-Gómez PA (1993) The multi-species surface-micelle model for surfactant adsorption and the BET equations. Colloid Polym. Sci. 271: 799–801.

    Google Scholar 

  • Haper KD, Bradshaw RA, Hartzell CR, Gurd FRN (1968) Comparison of myoglobins from harbor seal, porpoise, and sperm whale. I. Preparation and characterization. J. Biol. Chem. 243: 683–689.

    PubMed  Google Scholar 

  • Kelly JW (1996) Alternative conformations of amyloidogenic proteins govern their behavior. Curr. Opin. Struct. Biol. 6: 11–17.

    PubMed  Google Scholar 

  • Kuboi R, Yano K, Komasawa I (1994) Evaluation of surface properties and partitioning of proteins in aqueous two-phase extraction systems. Solv. Extr. Res. Dev. Japan. 1: 42–52.

    Google Scholar 

  • Raman B, Ramakrishna T, Rao CM (1996) Refolding of denatured and denatured/reduced lysozyme at high concentration. J. Biol. Chem. 271: 17067–17072.

    PubMed  Google Scholar 

  • Semisotnov GV, Rodionova NV, Razgulyaev OI, Uversky VN, Gripas AF, Gilmanshin RI (1991) Study of the molten globule intermediate state by hydrophobic fluorescent probe. Biopolymers 31: 119–128.

    PubMed  Google Scholar 

  • Stempfer G, Holl-Neugebauer B, Rodulph R (1996) Improved refolding of an immobilized fusion protein. Nat. Biotechnol. 14: 329–334.

    Google Scholar 

  • Stenberg E, Persson B, Roos H, Urbaniczky C (1991) Quantitative determination of surface concentration of protein with surface plasmon resonance using radiolabeled proteins. J. Coll. Int. Sci. 143: 513–526.

    Google Scholar 

  • Yamaguchi S, Mannen T, Zako T, Kamiya N, Nagamune T (2003) Measuring adsorption of a hydrophobic probe with a surface plasmon resonance sensor to monitor conformational changes in immobilized proteins. Biotechnol. Prog. 19: 1348–1354.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Chemistry and Biotechnology, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan

    Satoshi Yamaguchi, Teruhisa Mannen & Teruyuki Nagamune

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  1. Satoshi Yamaguchi
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  2. Teruhisa Mannen
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  3. Teruyuki Nagamune
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Yamaguchi, S., Mannen, T. & Nagamune, T. Evaluation of surface hydrophobicity of immobilized protein with a surface plasmon resonance sensor. Biotechnology Letters 26, 1081–1086 (2004). https://doi.org/10.1023/B:BILE.0000032969.32829.e3

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  • DOI: https://doi.org/10.1023/B:BILE.0000032969.32829.e3

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