- Physical Review Journals
All JournalsPhysics Magazine
Access Provided by Xinjiang University

Physical Review E

Export citation

Export citation

Choose format for download:

Download Citation
  • Access by Xinjiang University

Mechanism of protein binding to spherical polyelectrolyte brushes studied in situ using two-photon excitation fluorescence fluctuation spectroscopy

C. Czeslik* and R. Jansen

M. Ballauff* and A. Wittemann

C. A. Royer

E. Gratton and T. Hazlett

  • Physikalische Chemie I, Universität Dortmund, D-44221 Dortmund, Germany

  • Polymer-Institut, Universität Karlsruhe, D-76128 Karlsruhe, Germany

  • Centre de Biochimie Structurale, INSERM, F-34090 Montpellier, France

  • Laboratory for Fluorescence Dynamics, University of Illinois, Urbana, Illinois 61801, USA

  • *Corresponding author.
PDF

Phys. Rev. E 69, 021401 – Published 19 February, 2004

DOI: https://doi.org/10.1103/PhysRevE.69.021401

Abstract

We used two-photon excitation fluorescence fluctuation spectroscopy with photon counting histogram (PCH) analysis as a new tool to study the binding of globular proteins to colloidal particles in situ. Whereas fluorescence fluctuations are traditionally evaluated by calculating the autocorrelation function (fluorescence correlation spectroscopy), a complementary PCH analysis has been performed in this study which is advantageous when particle concentrations of a multicomponent system are of interest and the particles can be distinguished through particle brightness differences. The binding of two proteins, staphylococcal nuclease (SNase) and bovine serum albumin (BSA), to spherical polyelectrolyte brushes (SPB) was measured as a function of protein concentration and ionic strength of the solution at pH-values where SNase and BSA are positively and negatively charged, respectively. It has been found that SNase and BSA strongly bind to the SPB regardless of the protein charge. When the ionic strength of the solution is raised to 100 mM, the SPB become resistant to both proteins. These findings provide further evidence for a binding mechanism where the proteins are mainly driven to the SPB by the “counterion evaporation” force, while Coulomb interactions play a minor role. The results of this study characterize the potential of SPB as a new class of carrier particles for proteins whose use in biotechnological applications appears to be rewarding.

View figure in article

References (47)

  1. M. Fang, P. S. Grant, M. J. McShane, G. B. Sukhorukov, V. O. Golub, and Y. M. Lvov, Langmuir 18, 6338 (2002).
  2. F. Caruso and C. Schüler, Langmuir 16, 9595 (2000).
  3. F. Caruso and H. Möhwald, J. Am. Chem. Soc. 121, 6039 (1999).
  4. E. Donath, G. B. Sukhorukov, F. Caruso, S. A. Davis, and H. Möhwald, Angew. Chem. 110, 2323 (1998).
  5. F. Caruso, R. A. Caruso, and H. Möhwald, Science 282, 1111 (1998).
  6. C. M. Niemeyer, Angew. Chem., Int. Ed. Engl. 40, 4128 (2001).
  7. L. Yang and P. Alexandridis, Curr. Opin. Colloid Interface Sci. 5, 132 (2000).
  8. J. L. Ortega-Vinuesa and R. Hidalgo-Alvarez, Biotechnol. Bioeng. 47, 633 (1995).
  9. J. M. Peula, J. Puig, J. Serra, F. J. de las Nieves, and R. Hidalgo-Alvarez, Colloids Surf., A 92, 127 (1994).
  10. J.-Y. Yoon, H.-Y. Park, J.-H. Kim, and W.-S. Kim, J. Colloid Interface Sci. 177, 613 (1996).
  11. J. P. Santos, E. R. Welsh, B. P. Gaber, and A. Singh, Langmuir 17, 5361 (2001).
  12. A. Kondo, F. Murakami, and K. Higashitani, Biotechnol. Prog. 40, 889 (1992).
  13. W. Norde and J. P. Favier, Colloids Surf. 64, 87 (1992).
  14. P. Billsten, M. Wahlgren, T. Arnebrant, J. McGuire, and H. Elwing, J. Colloid Interface Sci. 175, 77 (1995).
  15. C. Czeslik, C. Royer, T. Hazlett, and W. Mantulin, Biophys. J. 84, 2533 (2003).
  16. P. Schwinté, V. Ball, B. Szalontai, Y. Haikel, J.-C. Voegel, and P. Schaaf, Biomacromolecules 3, 1135 (2002).
  17. P. Schwinté, J.-C. Voegel, C. Picart, Y. Haikel, P. Schaaf, and B. Szalontai, J. Phys. Chem. B 105, 11906 (2001).
  18. G. Decher, Science 277, 1232 (1997).
  19. A. Wittemann, B. Haupt, and M. Ballauff, Phys. Chem. Chem. Phys. 5, 1671 (2003).
  20. X. Guo and M. Ballauff, Phys. Rev. E 64, 051406 (2001).
  21. X. Guo and M. Ballauff, Langmuir 16, 8719 (2000).
  22. X. Guo, A. Weiss, and M. Ballauff, Macromolecules 32, 6043 (1999).
  23. G. Jackler, A. Wittemann, M. Ballauff, and C. Czeslik (unpublished).
  24. T. Neumann, B. Haupt, and M. Ballauff (unpublished).
  25. Physical Chemistry of Biological Interfaces, edited by A. Baszkin and W. Norde (Dekker, New York, 2000).
  26. Biopolymers at Interfaces, edited by M. Malmsten (Dekker, New York, 2003).
  27. C. A. Haynes, E. Sliwinsky, and W. Norde, J. Colloid Interface Sci. 164, 394 (1994).
  28. Y. Chen, J. D. Müller, P. T. C. So, and E. Gratton, Biophys. J. 77, 553 (1999).
  29. Y. Chen, J. D. Müller, K. M. Berland, and E. Gratton, Methods 19, 234 (1999).
  30. S. T. Hess, S. Huang, A. A. Heikal, and W. W. Webb, Biochemistry 41, 697 (2002).
  31. D. Magde, E. Elson, and W. W. Webb, Phys. Rev. Lett. 29, 705 (1972).
  32. D. C. Carter and J. X. Ho, Adv. Protein Chem. 45, 153 (1994).
  33. G. Ladam, P. Schaaf, G. Decher, J.-C. Voegel, and F. J. G. Cuisinier, Biomolec. Engineering 19, 273 (2002).
  34. S. Robinson and P. A. Williams, Langmuir 18, 8743 (2002).
  35. H. Seemann, R. Winter, and C. A. Royer, J. Mol. Biol. 307, 1091 (2001).
  36. Y. Mei, A. Wittemann, G. Sharma, M. Ballauff, T. Koch, H. Gliemann, J. Horbach, and T. Schimmel, Macromolecules 36, 3452 (2003).
  37. O. Krichevsky and G. Bonnet, Rep. Prog. Phys. 65, 251 (2002).
  38. K. M. Berland, P. T. So, and E. Gratton, Biophys. J. 68, 694 (1995).
  39. M. Placidi and S. Cannistraro, Europhys. Lett. 43, 476 (1998).
  40. J. D. Müller, Y. Chen, and E. Gratton, Biophys. J. 78, 474 (2000).
  41. G. Ladam, C. Gergely, B. Senger, G. Decher, J.-C. Voegel, P. Schaaf, and F. J. G. Cuisinier, Biomacromolecules 1, 674 (2000).
  42. V. P. Zhdanov and B. Kasemo, Langmuir 17, 5407 (2001).
  43. W. Norde, Macromol. Symp. 103, 5 (1996).
  44. C. M. Roth and A. M. Lenhoff, Langmuir 11, 3500 (1995).
  45. C. Fleck and H. H. von Grünberg, Phys. Rev. E 63, 061804 (2001).
  46. R. Hariharan, C. Biver, and W. B. Russel, Macromolecules 31, 7514 (1998).
  47. N. Dingenouts, R. Merkle, X. Guo, T. Narayanan, G. Goerigk, and M. Ballauff, J. Appl. Crystallogr. 36, 578 (2003).

Outline

Information

Phys. Rev. E 69, 021401– Published 19 February, 2004
  • Received 8 September 2003

© 2004 American Physical Society

Sign In to Your Journals Account

Filter

Filter

Article Lookup

Enter a citation