Abstract
The supra-molecular self-assembly of peptides and proteins is a process which underlies a range of normal and aberrant biological pathways in nature, but one which remains challenging to monitor in a quantitative way. We discuss the experimental details of an approach to this problem which involves the direct measurement in vitro of mass changes of the aggregates as new molecules attach to them. The required mass sensitivity can be achieved by the use of a quartz crystal transducer-based microbalance. The technique should be broadly applicable to the study of protein aggregation, as well as to the identification and characterisation of inhibitors and modulators of this process.
Key words
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
F. Chiti, C. M. Dobson, Protein misfolding, functional amyloid, and human disease, Annu Rev Biochem 75, 333 (2006).
C. M. Dobson, Protein folding and misfolding, Nature 426, 884 (2003).
M. B. Pepys, Pathogenesis, diagnosis and treatment of systemic amyloidosis, Phil. Trans. R. Soc. Lond. B 356, 203 (2001).
A. Aguzzi, Understanding the diversity of prions, Nat Cell Biol 6, 290 (2004).
D. M. Fowler, A. V. Koulov, W. E. Balch, J. W. Kelly, Functional amyloid–from bacteria to humans, Trends Biochem Sci 32, 217 (2007).
M. R. Chapman, L. S. Robinson, J. S. Pinkner, R. Roth, J. Heuser, M. Hammar, S. Normark, S. J. Hultgren, Role of Escherichia coli curli operons in directing amyloid fiber formation, Science 295, 851 (2002).
R. B. Wickner, [URE3] as an altered URE2 protein: evidence for a prion analog in Saccharomyces cerevisiae., Science 264, 566 (1994).
R. Krishnan, S. L. Lindquist, Structural insights into a yeast prion illuminate nucleation and strain diversity, Nature 435, 765 (2005).
M. Tanaka, S. R. Collins, B. H. Toyama, J. S. Weissman, The physical basis of how prion conformations determine strain phenotypes, Nature 442, 585 (2006).
C. M. Dobson, T. P. J. Knowles, M. E. Welland, UK patent application 0609382.7 (2006).
T. P. J. Knowles, W. Shu, G. L. Devlin, S. Meehan, S. Auer, C. M. Dobson, M. E. Welland, Kinetics and thermodynamics of amyloid formation from direct measurements of fluctuations in fibril mass, Proc Natl Acad Sci U S A 104, 10016 (2007).
D. A. White, A. K. Buell, C. M. Dobson, M. E. Welland, T. P. J. Knowles, Biosensor-based label-free assays of amyloid growth., FEBS Lett 583, 2587 (2009).
M. B. Hovgaard, M. Dong, D. E. Otzen, F. Besenbacher, Quartz crystal microbalance studies of multilayer glucagon fibrillation at the solid-liquid interface., Biophys J 93, 2162 (2007).
H. Okuno, K. Mori, T. Okada, Y. Yokoyama, H. Suzuki, Development of aggregation inhibitors for amyloid-beta peptides and their evaluation by quartz-crystal microbalance., Chem Biol Drug Des 69, 356 (2007).
J. A. Kotarek, K. C. Johnson, M. A. Moss, Quartz crystal microbalance analysis of growth kinetics for aggregation intermediates of the amyloid-beta protein., Anal Biochem (2008).
K. Hasegawa, K. Ono, M. Yamada, H. Naiki, Kinetic modeling and determination of reaction constants of Alzheimer’s beta-amyloid fibril extension and dissociation using surface plasmon resonance, Biochemistry 41, 13489 (2002).
A. K. Buell, G. G. Tartaglia, N. R. Birkett, C. A. Waudby, M. Vendruscolo, X. Salvatella, M. E. Welland, C. M. Dobson, T. P. J. Knowles, Position-dependent electrostatic protection against protein aggregation., Chembiochem 10, 1309 (2009).
D. A. White, A. K. Buell, T. P. J. Knowles, M. E. Welland, C. M. Dobson, Protein aggregation in crowded environments., J Am Chem Soc 132, 5170 (2010).
G. Sauerbrey, Verwendung von Schwingquarzen zur Wägung dünner Schichten und zur Mikrowägung, Z. Phys 155, 206 (1959).
T. Nomura, O. M, Frequency shifts of piezoelectric quartz crystals immersed in organic liquids, Anal. Chim. Acta 142, 281 (1981).
J. J. Kanazawa, J. G. Gordon, Frequency of a Quartz Microbalance in Contact with Liquid, Anal. Chem 57, 1771 (1985).
M. V. Voinova, M. Rodahl, M. Jonson, K. B, Viscoelastic Acoustic Responses of Layered Polymer Films at Fluid-Solid Interfaces: Continuum Mechanics Approach, Physica Scripta 59, 391 (1997).
S. S. Rogers, M. R. H. Krebs, E. H. C. Bromley, E. van der Linden, A. M. Donald, Optical microscopy of growing insulin amyloid spherulites on surfaces in vitro, Biophys J 90, 1043 (2006).
I. Reviakine, A. N. Morozov, F. F. Rosetti, Effects of finite crystal size in the quartz crystal microbalance with dissipation measurement system: Implications for data analysis, J. Appl. Phys 95, 7712 (2004).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Knowles, T.P.J., Devlin, G.L., Dobson, C.M., Welland, M.E. (2011). Probing Protein Aggregation with Quartz Crystal Microbalances. In: Hill, A., Barnham, K., Bottomley, S., Cappai, R. (eds) Protein Folding, Misfolding, and Disease. Methods in Molecular Biology, vol 752. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-223-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-60327-223-0_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-221-6
Online ISBN: 978-1-60327-223-0
eBook Packages: Springer Protocols