Simultaneous radial and wavelength analysis with the Optima XL-A analytical ultracentrifuge

  • P. Schuck
Conference paper
Part of the Progress in Colloid & Polymer Science book series (PROGCOLLOID, volume 94)


We exploit the possibility offered by the new analytical ultracentrifuge Beckman Optima XL-A to record wavelength profiles at different radial positions and radial profiles at different wavelengths in sedimentation equilibrium experiments to unravel complexes formed by entities with different absorption spectra. The mathematical analysis is described here in detail. With a little a priori knowledge about the absorption spectra it is possible to resolve a greater number of components than with conventional methods. This method also enables one to achieve a simultaneous detection of the components’ extinction profiles. In addition, an analysis of each component’s statistical accuracy is presented.

Key words

Analytical ultracentrifugation sedimentation equilibrium heterogeneous associations exponential fitting 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Shubert D, Schuck P (1991) Colloid Polym Sci 86:12–22Google Scholar
  2. 2.
    Hsu CS, Minton AP (1991) J Mol Recogn 4:93–104CrossRefGoogle Scholar
  3. 3.
    Varah JM (1985) SIAM J Sci Stat Comput 6:30–44CrossRefGoogle Scholar
  4. 4.
    Hanlon S, Lamers K, Lauterbach G, Johnson R, Schachman HK (1962) Arch Biochem Biophys 99:157–174CrossRefGoogle Scholar
  5. 5.
    Schachman HK, Gropper L, Hanlon S, Putney F (1962) Arch Biochem Biophys 99:175–190CrossRefGoogle Scholar
  6. 6.
    Osborne JC, Powell GM, Brewer HB (1980) Biochim Biophys Acta 619:559–571Google Scholar
  7. 7.
    Mulzer K, Kampmann L, Petrasch P, Schubert D (1990) Colloid Polym Sci 268:60–64CrossRefGoogle Scholar
  8. 8.
    Laue TM, Senear DF, Eaton S, Ross JBA (1993) Biochemistry 32:2469–2472CrossRefGoogle Scholar
  9. 9.
    Schuck P, Schubert D (1991) FEBS Lett 293:81–84CrossRefGoogle Scholar
  10. 13.
    Servillo L, Brewer HB, Osborne JC (1981) Biophys Chem 13:29–38CrossRefGoogle Scholar
  11. 11.
    Lewis MS (1993) Abstracts, VIII International Symposium on Ultracentrifugation, OsnabrückGoogle Scholar
  12. 12.
    Lakatos S, Minton P (1991) J Biol Chem 266:18707–18713Google Scholar
  13. 13.
    Pappert G, Schubert D (1983) Biochim Biophys Acta 730:32–40CrossRefGoogle Scholar
  14. 14.
    Passow H (1986) Rev Physiol Biochem Pharmacol 103:61–203Google Scholar
  15. 15.
    Johnson ML, Correia JJ, Yphantis DA, Halvorson HR (1981) Biophys J 36:575–588Google Scholar
  16. 16.
    Johnson ML, Lindsay MF (1992) Methods Enzymol 210:1–37CrossRefGoogle Scholar
  17. 17.
    Lawson CL, Hanson RJ (1974) Solving Least Squares Problems. Prentice-Hall, Englewood Cliffs, New JerseyGoogle Scholar
  18. 18.
    O'Leary DP, Rust BW (1986) SIAM J Sci Stat Comput 7:473–489CrossRefGoogle Scholar
  19. 19.
    Fujita H (1975) Foundations of Utracentrifugal Analysis. John Wiley & Sons, New YorkGoogle Scholar
  20. 20.
    Twomey S, Howell HB (1967) Appl Opt 6:2125–2131Google Scholar
  21. 21.
    Evans JW, Gragg WB, LeVeque RJ (1980) Math Comp 34:203–211CrossRefGoogle Scholar
  22. 22.
    Ruhe A (1980) SIAM J Sci Stat Comput 1:481–498CrossRefGoogle Scholar
  23. 23.
    Beechem JM (1992) Methods Enzymol 210:37–67Google Scholar
  24. 24.
    Kotaki A, Naoi M, Yagi K (1971) Biochim Biophys Acta 229:547–556Google Scholar
  25. 25.
    Dorst HJ, Schubert D (1979) Hoppe-Seyler's Z Physiol Chem 360:1605–1618Google Scholar
  26. 26.
    Casey JR, Lieberman DM, Reithmeier AF (1989) Methods Enzymol 173:494–512Google Scholar
  27. 27.
    Salhany JM (1990) Erythrocyte band 3 protein. CRC Press, Boca Raton, FloridaGoogle Scholar
  28. 28.
    Provencher SW (1982) Comput Phys Commun 27:213–227CrossRefGoogle Scholar
  29. 29.
    Lechner MD, Mächtle W (1991) Progr Colloid Polym Sci 86:62–69Google Scholar

Copyright information

© Dr. Dietrich Steinkopff Verlag GmbH & Co. KG 1994

Authors and Affiliations

  • P. Schuck
    • 1
  1. 1.Institut für Biophysik der J.W. Goethe-Universität FrankfurtFrankfurt/MainGermany

Personalised recommendations