Cooperative Assembly of Hsp70 Subdomain Clusters

Autor: Wright, Maya A, Aprile, Francesco A, Bellaiche, Mathias MJ, Michaels, Thomas CT, Müller, Thomas, Arosio, Paolo, Vendruscolo, Michele, Dobson, Christopher M, Knowles, Tuomas PJ
Přispěvatelé: Aprile, Francesco A [0000-0002-5040-4420], Arosio, Paolo [0000-0002-2740-1205], Vendruscolo, Michele [0000-0002-3616-1610], Knowles, Tuomas PJ [0000-0002-7879-0140], Apollo - University of Cambridge Repository
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Zdroj: Biochemistry
ISSN: 1520-4995
0006-2960
Popis: Many molecular chaperones exist as oligomeric complexes in their functional states, yet the physical determinants underlying such self-assembly behavior, as well as the role of oligomerization in the activity of molecular chaperones in inhibiting protein aggregation, have proven to be difficult to define. Here, we demonstrate direct measurements under native conditions of the changes in the average oligomer populations of a chaperone system as a function of concentration and time and thus determine the thermodynamic and kinetic parameters governing the self-assembly process. We access this self-assembly behavior in real time under native-like conditions by monitoring the changes in the micrometer-scale diffusion of the different complexes in time and space using a microfluidic platform. Using this approach, we find that the oligomerization mechanism of the Hsp70 subdomain occurs in a cooperative manner and involves structural constraints that limit the size of the species formed beyond the limits imposed by mass balance. These results illustrate the ability of microfluidic methods to probe polydisperse protein self-assembly in real time in solution and to shed light on the nature and dynamics of oligomerization processes.
Databáze: OpenAIRE
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