MODELING OF OLIGOMERIZATION AND FIBRILLOGENESIS OF BETA-2 MICROGLOBULIN MUTANT FORMS

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription or Fee Access

Abstract


A study has been performed in order to develop the approaches for therapy of amyloidosis using a model object — amyloidogenic protein human beta2-microglobulin (β2m). The aim of the study was to obtain protein variants with amino acid substitutions modulating fibrillogenic properties of β2m which potentially could inhibit fibrillogenesis of the wild-type protein. The study consisted of the two steps: a) computer modelling of β2m associative properties when introducing amino acid substitutions in the primary structure resulting in a change in the degree of molecular symmetry and b) experimental investigation of selected protein variants properties by appropriate genetic constructs design and obtaining the desired recombinant β2m variants. Based on computer calculations virtual variants with double substitutions at the N- and C-terminal regions of the protein molecule have been selected that could potentially provide alterations of monomer the associative properties. These variants have been obtained as recombinant products resulting from expression of genetic constructs specially designed. To obtain the construct with substitutions in the central region of the protein molecule an original double polymerase chain reaction (PCR) method has been developed yielding the intermediate extended primer — a β2m gene fragment with target substitutions which was used as a forward primer in the second PCR. It has been shown that recombinant β2m variants are capable to form fibrils.

Full Text

Restricted Access

About the authors

I V Arteeva

Institute of Experimental Medicine, NWB RAMS

Email: s.gestalt@gmail.com

V V Egorov

Research Institute of Influenza, MH RF; NRC «Kurchatov Institute», Petersburg Nuclear Physics Institute


A N Gorshkov

Research Institute of Influenza, MH RF


Y P Garmay

Research Institute of Influenza, MH RF; NRC «Kurchatov Institute», Petersburg Nuclear Physics Institute


T D Aleinikova

Institute of Experimental Medicine, NWB RAMS


M M Shavlovsky

Institute of Experimental Medicine, NWB RAMS


References

  1. Serpell L.C., Sunde M., Benson M.D. et al. The protofilament substructure of amyloid fibrils // J. of Molecular Biology. — 2000. — Vol. 300, № 5. — P. 1033-1039.
  2. Rochet J.C., Lansbury P.T. Jr. Amyloid fibrillogenesis: themes and variations // Current Opinion in Structural Biology. — 2000. — Vol. 10, № 1. — P. 60-68.
  3. Glenner G.G. Amyloid Deposits and Amyloidosis. The b-Fibrilloses // The New England J. of Medicine. — 1980. — Vol. 302, № 23. — P. 1283-1292.
  4. Merlini G., Bellotti V. Molecular Mechanisms of Amyloidosis // J. of Medicine. — 2003. — Vol. 349, № 6. — P. 583-596.
  5. Georgiades C.S., Neyman E.G., Barish M.A. et al. Amyloidosis: Review and Manifestations // RadioGraphics. — 2004. — Vol. 24, № 2. — P. 405-416.
  6. Hammarstrom P., Schneider F., Kelly J.W. Trans-Suppression of Misfolding in an Amyloid Disease // Science. — 2001. — Vol. 293, № 5539. — P. 2459-2462.
  7. McCutchen S.L., Lai Z., Miroy G.J. et al. Comparison of lethal and nonlethal transthyretin variants and their relationship to amyloid disease // Biochemistry. — 1995. — Vol. 34, № 41. — P. 13527-13536.
  8. Sebastiao M.P., Lamzin V., Saraiva M.J. et al. Transthyretin stability as a key factor in amyloidogenesis: X-ray analysis at atomic resolution // J. of Molecular Biology. — 2001. — Vol. 306, № 4. — P. 733-744.
  9. Ivanova M.I., Sawaya M.R., Gingery M. et al. An amyloid-forming segment of beta2-microglobulin suggests a molecular model for the fibril // PNAS. — 2004. — Vol. 101, № 293. — P. 10584-10589.

Statistics

Views

Abstract - 44

PDF (Russian) - 0

Cited-By


PlumX

Dimensions

Refbacks

  • There are currently no refbacks.

Copyright (c) 2013 Arteeva I.V., Egorov V.V., Gorshkov A.N., Garmay Y.P., Aleinikova T.D., Shavlovsky M.M.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies