Field of Science

The barrier to amyloid formation is kinetic, not thermodynamic

One of the questions I have pondered in the past is why the functional form of a protein should correspond to its most thermodynamically stable structure. Although this assumption is built into almost all experimental and theoretical studies of protein folding, it is not at all obvious since one may imagine other forms which could have improved stability. For instance, two protein forms may differ in the presence of a hydrogen bond or two. Based on the location and connectivity of these bonds, sometimes this slight rearrangement can cause a radical change in function, but there's no good reason why it should in the general case.

The answer however is most obvious in case of amyloid, that endlessly intriguing protein form that is implicated in so many devastating neurological disorders. Amyloid is a very stable state is often highly resistant to temperature, pH and high salt conditions. It's fair to ask how stable or unstable it is with respect to functional, soluble forms of the same protein.

To answer this question, a team led by Christopher Dobson who is a world expert on amyloid performed a series of thermodynamic measurements on a diverse group of proteins in which they measured the free energy differences between the soluble and the amyloid state. The proteins included everything from the Aß protein found in Alzheimer's disease to human lysozyme and insulin. The finding was that the free energy differences (ranging from about 3 kcal/mol to 6 kcal/mol) are not terribly dependent on the exact sequence, an observation which would be consistent with the striking recently uncovered fact that amyloid formation can be induced in almost any protein independent of its sequence. In fact the free energy difference seemed to depend more on the length and seemed to be optimal for a length of 100 residues for which the amyloid form was most stable. The difference also sharply tipped away from amyloid for increasing lengths.

This observation seems to suggest that one consequence of evolving larger proteins might be steer them away from the amyloid state and is consistent with the fact that almost all amyloid proteins have relatively short lengths (for instance, the Alzheimer's disease amyloid protein Aß has a length of roughly 40 residues). The propensity toward amyloid formation also depended on the concentration and the authors derived an limiting concentration beyond which amyloid formation would be rapid. This is again not surprising since the concentration-dependence of the process has also been demonstrated.

The real surprise came when they compared these limiting concentrations of the protein to the corresponding physiological concentrations of the same proteins in plasma. Remarkably, they found that in almost every case the physiological concentration was higher than that required to achieve amyloid formation. Thus
the observations clearly indicate that for many key proteins, the amyloid state is thermodynamically more stable than the native, functional state. To put it bluntly, many nicely folded and soluble proteins are actually metastable. Now, since native proteins don't constantly form amyloid and kill us all, it's clear that the barrier to amyloid formation must be kinetic. Intriguingly, the authors speculate that these barriers can be overcome when organisms are exposed to stress, mutations or aging.

This is a pretty intriguing study and seems to underscore the belief that at least for some proteins, the folded functional state is not the most stable. However in light of what we know about evolution, this should not be too surprising. Stability is just one of many factors to be optimized during natural selection and there is no reason to assume that evolution would always act to maximize this parameter at the cost of all others. It's worth always keeping in mind that evolution cannot afford to aim for the ideal but instead has to make do with what it has.

The other question in my mind is why in spite of these barriers existing in case of so many proteins like lysozyme, insulin etc. are they regularly overcome only in the case of Aß (1-42) and a select few others. Based on the speculation in the paper, this could be because these proteins are exposed to particularly harsh conditions that force them to climb past the kinetic barrier and settle into the amyloid valley of thermodynamic comfort and physiological woe.

Among many such conditions could very well be bacterial infections. A few years back I advanced a hypothesis about amyloid formation being a defense against viral and bacterial infection mediated through the production of free radicals. A kinetic barrier-surpassing mechanism of the kind speculated here might well be what allows these proteins to achieve the transition, killing the bacteria but ironically harming their owner in the process. In the context of the present study, I think there continue to be a lot of opportunities to investigate the possible infection-induced conversion of normal proteins to their amyloid form.

Hopefully someone will do the experiment.

Baldwin, A., Knowles, T., Tartaglia, G., Fitzpatrick, A., Devlin, G., Shammas, S., Waudby, C., Mossuto, M., Meehan, S., Gras, S., Christodoulou, J., Anthony-Cahill, S., Barker, P., Vendruscolo, M., & Dobson, C. (2011). Metastability of Native Proteins and the Phenomenon of Amyloid Formation Journal of the American Chemical Society DOI: 10.1021/ja2017703


  1. Hi Ash!

    Fascinating piece, and particularly boggling for a first year biochem student. The use of evolutionary analysis was greatly appreciated (stability as an under-optimized parameter, etc), but I have one question.

    You state " observation which would be consistent with the striking recently uncovered fact that amyloid formation can be induced in almost any protein independent of its sequence."

    As the cited paper is behind a paywall, I was unable to find the source of this undoubtedly awesome experiment.

    Is is available to the public, and if not, has it been covered in depth somewhere within the blogosphere?

    Thank you, and keep up the great work.

  2. Thanks. If you could send me an email at I would be happy to email you the papers.


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