An oligomeric equilibrium intermediate as the precursory nucleus of globular and fibrillar supramacromolecular assemblies in a PDZ domain.

Biophysical Journal
Javier Murciano-CallesJ C Martínez

Abstract

The equilibrium unfolding at neutral pH of the third PDZ domain of PSD95, as followed by DSC, is characterized by the presence of an equilibrium intermediate with clear signs of oligomerization. DLS and SEC measurements indicate that at 60-70 degrees C small oligomers populate, showing a typical beta-sheet far-UV CD spectrum. These intermediate species lead to the formation of rodlike particulates of approximately 12 nm, which remain in solution after 2 weeks incubation and grow until they adopt annular/spherical shapes of approximately 50 nm and protofibrils, which are subsequently fully transformed into fibrils. The fibrils can also disaggregate after the addition of 1:1 buffer dilution followed by cooling to room temperature, thus returning to the initial monomeric state. Growth kinetics, as shown by ThT and ANS fluorescence, show that the organization of the different supramacromolecular structures comes from a common nucleation unit, the small oligomers, which organize themselves before reaching the incubation temperature of 60 degrees C. Our experiments point toward the existence of a well-defined reversible, stepwise, and downhill organization of the processes involved in the association-dissociation of the intermediate....Continue Reading

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Citations

Oct 2, 2012·Archives of Biochemistry and Biophysics·Christopher M Johnson
Jun 28, 2012·Biophysical Journal·Gabriela María TorchioMauricio Pablo Sica
May 26, 2012·The Journal of Physical Chemistry. B·Montserrat Andujar-SanchezJose C Martinez
Nov 20, 2019·Journal of Molecular Recognition : JMR·Javier Murciano-CallesJose C Martinez
Apr 17, 2019·Acta Crystallographica. Section D, Structural Biology·Ana Camara-ArtigasJose C Martínez
Jul 31, 2020·Life·Javier Murciano-Calles
Nov 19, 2020·Nature Communications·Olga BozovicPeter Hamm
Jan 6, 2018·The Journal of Physical Chemistry. B·Zhenxing Liu, D Thirumalai

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