Zinc-Dependent Oligomerization of Thermus thermophilus Trigger Factor Chaperone

Thermus thermophilus trigger factor (TtTF) is a zinc-dependent molecular chaperone whose folding-arrest activity is regulated by Zn²⁺. However, little is known about the mechanism of zinc-dependent regulation of the TtTF activity. Here we exploit in vitro biophysical experiments to investigate zinc-binding, the oligomeric state, the secondary structure, and the thermal stability of TtTF in the absence and presence of Zn²⁺. The data show that full-length TtTF binds Zn²⁺, but the isolated domains and tandem domains of TtTF do not bind to Zn²⁺. Furthermore, circular dichroism (CD) and nuclear magnetic resonance (NMR) spectra suggested that Zn²⁺-binding induces the partial structural changes of TtTF, and size exclusion chromatography-multi-angle light scattering (SEC-MALS) showed that Zn²⁺ promotes TtTF oligomerization. Given the previous work showing that the activity regulation of E. coli trigger factor is accompanied by oligomerization, the data suggest that TtTF exploits zinc ions to induce the structural change coupled with the oligomerization to assemble the client-binding site, thereby effectively preventing proteins from misfolding in the thermal environment.