  The cornerstone of structural biology is the unique relationship between
protein sequence and the 3D structure at equilibrium. Although intrinsically
disordered proteins (IDPs) do not fold into a specific 3D structure, breaking
this paradigm, some IDPs exhibit large-scale organization, such as
liquid-liquid phase separation. In such cases, the structural plasticity has
the potential to form numerous self-assembled structures out of thermal
equilibrium. Here, we report that high-temperature incubation time is a
defining parameter for micro and nanoscale self-assembly of resilin-like IDPs.
Interestingly, high-resolution scanning electron microscopy micrographs reveal
that an extended incubation time leads to the formation of micron-size rods and
ellipsoids that depend on the amino acid sequence. More surprisingly, a
prolonged incubation time also induces amino acid composition-dependent
formation of short-range nanoscale order, such as periodic lamellar
nanostructures. We, therefore, suggest that regulating the period of
high-temperature incubation, in the one-phase regime, can serve as a unique
method of controlling the hierarchical self-assembly mechanism of structurally
disordered proteins.
