Kinetics and thermodynamics of amyloid formation from direct measurements of fluctuations in fibril mass

被引:175
作者
Knowles, Tuomas P. J.
Shu, Wenmiao
Devlin, Glyn L.
Meehan, Sarah
Auer, Stefan
Dobson, Christopher M.
Welland, Mark E.
机构
[1] Univ Cambridge, Nanosci Ctr, Cambridge CB3 0FF, England
[2] Univ Cambridge, Cavendish Lab, Cambridge CB3 0HE, England
[3] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England
基金
英国惠康基金;
关键词
protein aggregation; quartz crystal microbalance; biosensors;
D O I
10.1073/pnas.0610659104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Aggregation of proteins and peptides is a widespread and much-studied problem, with serious implications in contexts ranging from biotechnology to human disease. An understanding of the proliferation of such aggregates under specific conditions requires a quantitative knowledge of the kinetics and thermodynamics of their formation; measurements that to date have remained elusive. Here, we show that precise determination of the growth rates of ordered protein aggregates such as amyloid fibrils can be achieved through real-time monitoring, using a quartz crystal oscillator, of the changes in the numbers of molecules in the fibrils from variations in their masses. We show further that this approach allows the effect of other molecular species on fibril growth to be characterized quantitatively. This method is widely applicable, and we illustrate its power by exploring the free-energy landscape associated with the conversion of the protein insulin to its amyloid form and elucidate the role of a chemical chaperone and a small heat shock protein in inhibiting the aggregation reaction.
引用
收藏
页码:10016 / 10021
页数:6
相关论文
共 47 条
[1]   Games played by rogue proteins in prion disorders and Alzheimer's disease [J].
Aguzzi, A ;
Haass, C .
SCIENCE, 2003, 302 (5646) :814-818
[2]   Partially folded intermediates in insulin fibrillation [J].
Ahmad, A ;
Millett, IS ;
Doniach, S ;
Uversky, VN ;
Fink, AL .
BIOCHEMISTRY, 2003, 42 (39) :11404-11416
[3]   Direct observation of Aβ amyloid fibril growth and inhibition [J].
Ban, T ;
Hoshino, M ;
Takahashi, S ;
Hamada, D ;
Hasegawa, K ;
Naiki, H ;
Goto, Y .
JOURNAL OF MOLECULAR BIOLOGY, 2004, 344 (03) :757-767
[4]   Formation of insulin amyloid fibrils followed by FTIR simultaneously with CD and electron microscopy [J].
Bouchard, M ;
Zurdo, J ;
Nettleton, EJ ;
Dobson, CM ;
Robinson, CV .
PROTEIN SCIENCE, 2000, 9 (10) :1960-1967
[5]   Therapeutic approaches to protein-misfolding diseases [J].
Cohen, FE ;
Kelly, JW .
NATURE, 2003, 426 (6968) :905-909
[6]   Mechanism of prion propagation: Amyloid growth occurs by monomer addition [J].
Collins, SR ;
Douglass, A ;
Vale, RD ;
Weissman, JS .
PLOS BIOLOGY, 2004, 2 (10) :1582-1590
[7]  
Demaimay R, 1998, J NEUROCHEM, V71, P2534
[8]   Prevention of polymerization of M and Z α1-antitrypsin (α1-AT) with trimethylamine N-oxide -: Implications for the treatment of α1-AT deficiency [J].
Devlin, GL ;
Parfrey, H ;
Tew, DJ ;
Lomas, DA ;
Bottomley, SP .
AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY, 2001, 24 (06) :727-732
[9]   Protein folding and misfolding [J].
Dobson, CM .
NATURE, 2003, 426 (6968) :884-890
[10]  
DOBSON CM, 2006, Patent No. 06093827