Abstract
Amyloid fibril formation is a phenomenon common to many proteins and peptides associated with numerous conformational diseases. To clarify the mechanism of fibril formation and to create inhibitors, real-time monitoring of fibril growth is essential. This chapter describes a method to visualize amyloid fibril growth in real time at the single fibril level. This approach uses total internal reflection fluorescence microscopy (TIRFM) combined with the binding of thioflavin T, an amyloidspecifi c fluorescence dye. The method enables an exact analysis of the rate of growth of individual fibrils. One of the advantages of TIRFM is that only amyloid fibrils lying in parallel with the slide glass surface were observed, so that one can obtain the exact length of fibrils. This method is of particular importance for the analysis of rapid fibrillation kinetics, providing unique information crucial for the elucidation of the molecular mechanisms of amyloid fibril formation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Ban, T., Hamada, D., Hasegawa, H., Naiki, H., and Goto, Y. (2003). Direct observation of amyloid fibril growth monitored by thioflavin T fluorescence. J. Biol. Chem. 278:16462–16465.
Bjokman, P.J., Saper, M.A., Samraoui, B., Bennett, W.S., Strominger, J.L., and Willy, D.C. (1987). Structure of the human class I histocompatibility antigen, HLA-A2. Nature 329:506–512.
Chiba, T., Hagihara, Y., Higurashi, T., Hasegawa, K., Naiki, H., and Goto, Y. (2003). Amyloid fibril formation in the context of full-length protein: effects of proline mutations on the amyloid fibril formation of β2-microglobulin. J. Biol. Chem. 278:47016–47024.
Depace, A.H., and Weissman, J. S. (2002). Origins and kinetic consequences of diversity in Sup35 yeast prion fibrils. Nat. Struct. Biol. 9:389–396.
Dobson, C.M. (2003). Protein folding and misfolding. Nature 426:884–889.
Funatsu, T., Harada, Y., Tokunaga, M., Saito, K., and Yanagida, T. (1995). Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution. Nature 374:555–559.
Gejyo, F., Yamada, T., Odani, S., Nakagawa, Y., Arakawa, M., Kunitomo, T., Kataoka, H., Suzuki, M., Nirasawa, Y., Shirahama, T., Cohen, A.S., and Schmid, K. (1985). A new form of amyloid protein associated with chronic hemodialysis was identified as β2-microglobulin. Biochem. Biophys. Res. Commun. 129:701–706.
Goldsbury, C., Kistler, J., Aebi, U., Arvinte, T., and Cooper, G.J. (1999). Watching amyloid fibrils grow by time-lapse atomic force microscopy. J. Mol. Biol. 285:33–39.
Green, J.D., Goldsbury, C., Kistler. J., Cooper. G.J.S., and Aebi, U. (2004). Human amylin oligomer growth and fibril elongation define two distinct phases in amyloid formation. J. Biol. Chem. 279:12206–12212.
Hardy, J.A., and Higgins, G.A. (1992). Alzheimer’s disease: the amyloid cascade hypothesis. Science 256:184–185.
Hardy, J., and Selkoe, D.J. (2002). The amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics. Science 297:353–356.
Hasegawa, K., Yamaguchi, I., Omata, S., Gejyo, F., and Naiki, H. (1999). Interaction between Aβ (1-42) and Aβ(1-40) in Alzheimer’s β-amyloid fibril formation in vitro. Biochemistry 38:15514–15521.
Hasegawa, K., Ono, K., Yamada, M., and Naiki, H. (2002). Kinetic modeling and determination of reaction constants of Alzheimer’s β-amyloid fibril extension and dissociation using surface plasmon resonance. Biochemistry 41:13489–13498.
Häggqvist, B., Näslund, J., Sletten, K., Westermark, G.T., Mucchiano, G., Tjernberg, L.O., Nordstedt, C., Engström, U., and Westermark, P. (1999). Medin: an integral fragment of aortic smooth muscle cell-produced lactadherin forms the most common human amyloid. Proc. Natl. Acad. Sci. USA 96:8669–8674.
Hoyer, W., Cherny, D., Subramaniam, V., and Jovin T.M. (2004). Rapid self-assembly of α-synuclein observed by in situ atomic force microscopy. J. Mol. Biol. 340:127–139.
Inoue, Y., Kishimoto, A., Hirao, J., Yoshida, M., and Taguchi, H. (2001). Strong growth polarity of yeast prion fiber revealed by single fiber imaging. J. Biol. Chem. 276:35227–35230.
Ionescu-Zanetti, C., Khurana, R., Gillespie, J.R., Petrick, J.S., Trabachino, L.C., Minert, L.J., Carter, S.A., and Fink, A.L. (1999). Monitoring the assembly of Ig light-chain amyloid fibrils by atomic force microscopy. Proc. Natl. Acad. Sci. USA 96:13175–13179.
Ippel, J.H., Olofsson, A., Schleucher, J.S., Lundgren, E., and Wijmenga, S.S. (2002). Probing solvent accessibility of amyloid fibrils by solution NMR spectroscopy. Proc. Natl. Acad. Sci. USA 99:8648–8653.
Kad, N.M., Myers, S.L., Smith, D.P., Smith, D.A., Radford, S.E., and Thomson, N.H. (2003). Hierarchical assembly of β2-microglobulin amyloid in vitro revealed by atomic force microscopy. J. Mol. Biol. 330:785–797.
Katou, H., Kanno, T., Hoshino, M., Hagihara, Y., Tanaka, H., Kawai, T., Hasegawa, K., Naiki, H., and Goto, Y. (2002). The role of disulfide bond in the amyloidogenic state of β2-microglobulin studied by heteronuclear NMR. Protein Sci. 11:2219–2229.
Kozhukh, G.V., Hagihara, Y., Kawakami, T., Hasegawa, K., Naiki, H., and Goto, Y. (2002). Investigation of a peptide responsible for amyloid fibril formation of β2-microglobulin by Acromobacter protease I. J. Biol. Chem. 277:1310–1315.
Mattson M.P. (2004). Pathways towards and away from Alzheimer’s disease. Nature 430:631–639.
Naili, H., and Gejyo, F. (1999). Kinetic analysis of amyloid fibril formation. Methods Enzymol. 309:305–318.
Naiki, H., Higuchi, K., Hosokawa, M., and Takeda, T. (1989). Fluorometric determination of amyloid fibrils in vitro using the fluorescent dye, thioflavin T. Anal. Biochem. 177:244–249.
Naiki, H., Hashimoto, N., Suzuki, S., Kimura, H., Nakakuki, K., and Gejyo, F. (1997). Establishment of a kinetic model of dialysis-related amyloid fibril extension in vitro. Amyloid 4:223–232.
Ohhashi, Y., Hagihara, Y., Kozhukh, G., Hoshino, M., Hasegawa, K., Yamaguchi, I., Naiki, H., and Goto, Y. (2002). The intrachain disulfide bond of β2-microglobulin is not essential for the immunoglobulin fold at neutral pH, but is essential for amyloid fibril formation at acidic pH. J. Biochem. 131:45–52.
Scheibel, T., Kowal, A.S., Bloom, J.D., and Lindquist, S.L. (2001). Bidirectional amyloid fiber growth for a yeast prion determinant. Curr. Biol. 11:366–369.
Wazawa, T., Ishii, Y., Funatsu, T., and Yanagida, T. (2000). Spectral fluctuation of a single fluorophore conjugated to a protein molecule. Biophys. J. 78:1561–1569.
Yamasaki, R., Hoshino, M., Wazawa, T., Ishii, Y., Yanagida, T., Kawata, Y. Higurashi, T., Sakai, K., Nagai, J., and Goto, Y. (1999). Single molecular observation of the interaction of GroEL with substrate proteins. J. Mol. Biol. 292:965–972.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Springer Science+Business Media, Inc.
About this chapter
Cite this chapter
Ban, T., Goto, Y. (2006). Direct Observation of Amyloid Fibril Growth Monitored by Total Internal Reflection Fluorescence Microscopy. In: Uversky, V.N., Fink, A.L. (eds) Protein Misfolding, Aggregation, and Conformational Diseases. Protein Reviews, vol 4. Springer, Boston, MA. https://doi.org/10.1007/0-387-25919-8_17
Download citation
DOI: https://doi.org/10.1007/0-387-25919-8_17
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-387-25918-5
Online ISBN: 978-0-387-25919-2
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)