Biochemistry (Moscow)

, Volume 78, Issue 4, pp 368–376 | Cite as

Role of cis- and trans-interactions in manifestations of amyloidogenic properties of variable domains of Bence-Jones proteins TIM and LUS

  • V. M. TishchenkoEmail author


Intact Bence-Jones proteins TIM and LUS under simulated physiological conditions (10 mM phosphate buffer, pH 7.0, 100 mM NaCl, 37°C) did not display amyloidogenic properties. However, their isolated variable domains exhibit these qualities in full measure. Therefore, both intact proteins and their variable domains were studied using a complex of physical methods (scanning microcalorimetry, analytical centrifugation, optics) that allowed us to assess the stability of their tertiary and quaternary structures. The experimentally obtained thermodynamic functions indicated that the stability of iso-lated variable domains of TIM and LUS was comparable to the stability of similar domains in amyloidogenic proteins described earlier. However, inside the whole protein their stability was comparable to the stability of VL domains of ordinary Bence-Jones proteins. The decreased stability of the isolated variable domains of TIM and LUS was shown to be due both to weak interactions between a pair of variable domains (trans -interaction) and to a natural lack of interaction with the con-stant domains (cis-interaction).

Key words

Bence-Jones proteins amyloid fibrils variable domains stabilization free energy interdomain interactions 



light chain constant domains


bis(N-hydroxysuccinimide suberate) ester


a fragment with variable domains of the light chain cross-linked with the corresponding reagent

Fb-subunit (fragment)

a structure formed by a pair of constant domains CL-CL


fluorescein isothio-cyanate

Fv-subunit (fragment)

a structure formed by a pair of variable domains VL-VL


FITC-labeled variable (constant) domain


immunoglobulins of G class


light chain variable domains


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  1. 1.
    Falk, R. H., Comenzo, R. L., and Skinner, M. (1997) N. Engl. J. Med., 337, 898–909.PubMedCrossRefGoogle Scholar
  2. 2.
    Huff, M. E., Balch, W. E., and Kelly, J. W. (2003) Curr. Opin. Struct. Biol., 13, 674–682.PubMedCrossRefGoogle Scholar
  3. 3.
    Pozzi, C., and Locatelli, F. (2002) Semin. Nephrol., 22, 319–330.PubMedGoogle Scholar
  4. 4.
    Helms, L. R., and Wetzel, R. (1995) Protein Sci., 4, 2073–2081.PubMedCrossRefGoogle Scholar
  5. 5.
    Wall, J., Schell, M., Murphy, C., Hrncic, R., Stevens, F. J., and Solomon, A. (1999) Biochemistry, 38, 14101–14108.PubMedCrossRefGoogle Scholar
  6. 6.
    Raffen, R., Dieckman, L. J., Szpunar, M., Wanschl, C., Rokkuluri, P. R., Dave, R., Wilkins, Stevens, P., Cai, X., Schiffer, M., and Stevens, F. J. (1999) Protein Sci., 8, 509–517.PubMedCrossRefGoogle Scholar
  7. 7.
    Kim, Y., Wall, J. S., Meyer, J., Murphy, C., Randolph, T. W., Manning, M., Solomon, A., and Carpenter, J. F. (2000) J. Biol. Chem., 275, 1570–1574.PubMedCrossRefGoogle Scholar
  8. 8.
    Solomon, A., Weiss, D. T., Murphy, C. L., Hrncic, R., Wall, J. S., and Schell, M. (1998) Proc. Natl. Acad. Sci. USA, 95, 9547–9551.PubMedCrossRefGoogle Scholar
  9. 9.
    Tishchenko, V. M., Khristoforov, V. S., and Bliznyukov, O. P. (2009) Mol. Biol., 43, 148–156.CrossRefGoogle Scholar
  10. 10.
    Tishchenko, V. M. (2011) Mol. Biol., 45, 1055–1064.CrossRefGoogle Scholar
  11. 11.
    Epp, O., Lattman, E. E., Schiffer, M., Huber, R., and Palm, W. (1975) Biochemistry, 14, 943–952.CrossRefGoogle Scholar
  12. 12.
    Chang, C. H., Short, M. T., Westholm, F. A., Stevens, F. J., Wang, B. C., Furey, W., Jr., Solomon, A., and Schiffer, M. (1985) Biochemistry, 24, 4890–4897.PubMedCrossRefGoogle Scholar
  13. 13.
    Schiffer, M., Chang, C.-H., Naik, V. M., and Stevens, F. J. (1988) J. Mol. Biol., 203, 799–802.PubMedCrossRefGoogle Scholar
  14. 14.
    Miller, S. (1990) J. Mol. Biol., 216, 965–973.PubMedCrossRefGoogle Scholar
  15. 15.
    Bliznyukov, O. P., Kozmin, L. D., Vysotskaya, L. L., Golenkov, A. K., Tishchenko, V. M., Samoilovich, M. P., and Klimovich, V. P. (2005) Biochemistry (Moscow), 70, 458–466.CrossRefGoogle Scholar
  16. 16.
    Laemmli, U. K. (1970) Nature, 227, 680–685.PubMedCrossRefGoogle Scholar
  17. 17.
    Laurell, C. B. (1965) Analyt. Biochem., 10, 358–361.PubMedCrossRefGoogle Scholar
  18. 18.
    Zav’yalov, V. P., Troitsky, G. V., Khechinashvili, N. N., and Privalov, P. L. (1977) Biochim. Biophys. Acta, 492, 102–111.PubMedCrossRefGoogle Scholar
  19. 19.
    Tishchenko, V. M. (2001) Biochemistry (Moscow), 66, 1352–1355.CrossRefGoogle Scholar
  20. 20.
    Tischenko, V. M., Abramov, V. M., and Zav’yalov, V. P. (1998) Biochemistry, 37, 5576–5581.PubMedCrossRefGoogle Scholar
  21. 21.
    Tischenko, V. M., and Zav’yalov, V. P. (2002) Immunol. Lett., 84, 241–245.PubMedCrossRefGoogle Scholar
  22. 22.
    Eulitz, M., Chang, L.-Y., Zirkel, C., Schell, M., Weiss, D. T., and Solomon, A. (1995) J. Immunol., 154, 3256–3265.PubMedGoogle Scholar
  23. 23.
    Wall, J., Murphy, C. L., and Solomon, A. (1999) Methods Enzymol., 309, 204–216.PubMedCrossRefGoogle Scholar
  24. 24.
    Yphantis, D. A. (1964) Biochemistry, 3, 297–317.PubMedCrossRefGoogle Scholar
  25. 25.
    Van Holde, K. E., and Baldwin, R. L. (1958) J. Phys. Chem., 62, 734–743.CrossRefGoogle Scholar
  26. 26.
    Bowen, T. (1971) in An Introduction to Ultracentrifugation (Degly, S., ed.) Wiley-Interscience, London-N.-Y.-Sydney-Toronto, pp. 107–108.Google Scholar
  27. 27.
    Tishchenko, V. M. (2000) Mol. Biol., 34, 116–122.Google Scholar
  28. 28.
    Privalov, P. L., and Potekhin, S. A. (1986) Methods Enzymol., 131, 4–51.PubMedCrossRefGoogle Scholar
  29. 29.
    Wetzel, R. (1997) Adv. Prot. Chem., 50, 183–242.CrossRefGoogle Scholar
  30. 30.
    Tishchenko, V. M. (2012) Biofizika, in press.Google Scholar
  31. 31.
    Filimonov, V. V., and Rogov, V. V. (1996) J. Mol. Biol., 255, 767–777.PubMedCrossRefGoogle Scholar
  32. 32.
    Potekhin, S. A., and Kovrigin, E. L. (1998) Biofizika, 43, 223–232.PubMedGoogle Scholar
  33. 33.
    Potekhin, S. A., Loseva, O. I., Tiktopulo, E. I., and Dobritsa, A. P. (1999) Biochemistry, 38, 4121–4127.PubMedCrossRefGoogle Scholar
  34. 34.
    Tischenko, V. M., Zav’yalov, V. P., Medgyesi, G. A., Potekhin, S. A., and Privalov, P. L. (1982) Eur. J. Biochem., 126, 517–521.PubMedCrossRefGoogle Scholar
  35. 35.
    Zav’yalov, V. P., and Tishchenko, V. M. (1991) Scand. J. Immunol., 33, 755–762.PubMedCrossRefGoogle Scholar
  36. 36.
    Tischenko, V. M., and Zav’yalov, V. P. (2003) Immunol. Lett., 86, 281–285.PubMedCrossRefGoogle Scholar
  37. 37.
    Privalov, P. L. (1979) Adv. Prot. Chem., 33, 167–241.CrossRefGoogle Scholar
  38. 38.
    Privalov, P. L. (1982) Adv. Prot. Chem., 35, 1–104.CrossRefGoogle Scholar
  39. 39.
    Privalov, P. L., and Makhatadze, G. I. (1992) J. Mol. Biol., 224, 715–723.PubMedCrossRefGoogle Scholar
  40. 40.
    Tischenko, V. M. (1999) in Abst. XIth Int. Conf. on Small-Angle Scattering, Brookhaven National Laboratory, N. Y., p. 93.Google Scholar
  41. 41.
    Tishchenko, V. M. (2013) Mol. Biol., in press.Google Scholar
  42. 42.
    Del Pozo Yauner, L., Ortiz, E., Sanchez, R., Sanchez-Lopez, R., Guereca, L., Murphy, C. L., Allen, A., Wall, J. S., Fernandez-Velasco, D. A., Solomon, A., and Becerril, B. (2008) Proteins, 72, 684–692.PubMedCrossRefGoogle Scholar
  43. 43.
    Rowe, E. S., and Tanford, C. (1970) Biochemistry, 24, 4822–4827.Google Scholar
  44. 44.
    Goto, Y., Ichimura, N., and Hamaguchi, K. (1988) Biochemistry, 27, 1670–1677.PubMedCrossRefGoogle Scholar
  45. 45.
    Pace, C. N., Shirley, B. A., and Thomson, J. A. (1989) in Protein Structure: A Practical Approach (Creighton, T. E., ed.) IRL Press, Oxford, pp. 311–330.Google Scholar
  46. 46.
    Blancas-Mejia, L. M., Tellez, L. A., del Pozo-Yauner, L., Becerril, B., Sanchez-Ruiz, J. M., and Fernandez-Velasco, D. A. (2009) J. Mol. Biol., 386, 1153–1166.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2013

Authors and Affiliations

  1. 1.Institute of Theoretical and Experimental BiophysicsRussian Academy of SciencesPushchino, Moscow RegionRussia

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