Abstract: Oligomers are commonly observed intermediates at the initial stages of amyloid fibril formation. They are toxic to neurons and cause decrease in neural transmission and long-term potentiation. We describe an in vitro study of the initial steps in amyloid fibril formation by human stefin B, which proved to be a good model system. Due to relative stability of the initial oligomers of stefin B, electrospray ionization mass spectrometry (ESI MS) could be applied in addition to size exclusion chromatography (SEC). These two techniques enabled us to separate and detect distinguished oligomers from the monomers: dimers, trimers, tetramers, up to decamers. The amyloid fibril formation process was followed at different pH and temperatures, including such conditions where the process was slow enough to detect the initial oligomeric species at the very beginning of the lag phase and those at the end of the lag phase. Taking into account the results of the lower-order oligomers transformations early in the process, we were able to propose an improved model for the stefin B fibril formation.
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Taler-Verčič, A.; Kirsipuu, T.; Friedemann, M.; Noormägi, A.; Polajnar, M.; Smirnova, J.; Žnidarič, M.T.; Žganec, M.; Škarabot, M.; Vilfan, A.; Staniforth, R.A.; Palumaa, P.; Žerovnik, E. The Role of Initial Oligomers in Amyloid Fibril Formation by Human Stefin B. Int. J. Mol. Sci. 2013, 14, 18362-18384.
Taler-Verčič A, Kirsipuu T, Friedemann M, Noormägi A, Polajnar M, Smirnova J, Žnidarič MT, Žganec M, Škarabot M, Vilfan A, Staniforth RA, Palumaa P, Žerovnik E. The Role of Initial Oligomers in Amyloid Fibril Formation by Human Stefin B. International Journal of Molecular Sciences. 2013; 14(9):18362-18384.
Taler-Verčič, Ajda; Kirsipuu, Tiina; Friedemann, Merlin; Noormägi, Andra; Polajnar, Mira; Smirnova, Julia; Žnidarič, Magda T.; Žganec, Matjaž; Škarabot, Miha; Vilfan, Andrej; Staniforth, Rosemary A.; Palumaa, Peep; Žerovnik, Eva. 2013. "The Role of Initial Oligomers in Amyloid Fibril Formation by Human Stefin B." Int. J. Mol. Sci. 14, no. 9: 18362-18384.