Ulf Olsson

The chaperone DNAJB6b (JB6) plays important roles in increasing amyloid protein solubility and inhibiting amyloid fibril formation, a causative factor for neurodegenerative diseases like Alzheimer's and Parkinson's disease. Insights into the biophysical properties of JB6, including its structure, self-assembly and stability towards denaturation, may enhance the understanding of the physicochemical basis of chaperone action. However, many of the biophysical properties of the chaperone remain elusive. Here, we investigated the structure and stability of JB6 and its domains towards thermal and chemical denaturation using Fourier transform infrared and circular dichroism spectroscopy to examine the thermodynamic properties. Both domains act as independent folding units. We find that the N-terminal domain (NTD) of JB6 is more stable than its C-terminal domain (CTD). Both domains are stabilized in the context of the full-length protein. The intact protein unfolds in a step-wise manner when subjected to a denaturing agent with the CTD unfolding at a lower denaturant concentration than the NTD. The combination of thermal and chemical denaturation followed by differential scanning fluorimetry revealed the enthalpy changes (22.6 and 26.4 kJ mol⁻¹) and heat capacity changes (2.8 and 3.0 kJ/(mol*K)) upon denaturation of NTD alone and of NTD within the full-length protein, respectively. The understanding of JB6's biophysical properties complements the increasing amount of data on JB6's interactions with client proteins, paving the way for further investigation of the mechanism of its cellular function.