Folding and Misfolding Dynamics of Irisin Protein Revealed by Single-Molecule Magnetic Tweezers

Irisin, a fibronectin III protein secreted by muscles during physical exercise, plays a significant role in the browning of white fat and cell adhesion, highlighting the importance of its conformational transitions. In this study, we investigated the folding and unfolding dynamics of a single irisin domain using a single-molecule manipulation technique known as magnetic tweezers. In addition to the native state, irisin can also fold transiently into a misfolded state. We determined the folding free energies of the native and misfolded states as well as their force-dependent folding and unfolding rates. The free energy of the misfolded state is higher than that of the unfolded state, and the misfolded state has a homogeneous force-dependent unfolding rate. The stable native state demonstrates heterogeneous unfolding rates that are within ∼1 order of magnitude. Via comparison with the well-studied 10th fibronectin III domain that has a partially folded intermediate state, our study demonstrates that proteins with similar structure can have distinct folding pathways.