Modulating Hierarchical Self-Assembly In Thermoresponsive Intrinsically Disordered Proteins Through High-Temperature Incubation Time

TL;DR

This study demonstrates that high-temperature incubation time critically influences the hierarchical self-assembly of intrinsically disordered proteins, enabling control over their micro- and nanoscale structures.

Contribution

It reveals that incubation time at high temperature modulates the self-assembly pathways and structures of IDPs, providing a new method for controlling protein organization.

Findings

Extended incubation leads to micron-sized rods and ellipsoids.

Incubation time induces amino acid-dependent nanoscale lamellar structures.

High-temperature incubation time can be used to regulate hierarchical self-assembly.

Abstract

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.