Structural basis and mechanism of the unfolding-induced activation of an acid response chaperone HdeA

Abstract

The role of protein structural disorder in biological functions is gaining increasing interests in the past decade. The bacterial acid-resistant chaperone HdeA belongs to a group of \"conditionally disordered\" protein that is activated via an order-to-disorder transition. However, the mechanism for unfolding-induced activation remains unclear due to the lack of experimental information on the unfolded state conformation and the chaperone-client interactions. Here we use advanced solution NMR methods to characterize the activated state conformation of HdeA under acidic condition and identify the client binding sites. The activated HdeA becomes largely disordered and exposes two essential hydrophobic patches of residues for client interactions. The pH-dependent chemical exchange saturation transfer (CEST) result identifies three acid-sensitive regions that act as structural locks during the activation process, revealing a multi-step activation mechanism of HdeA chaperone function at atomic level. The results highlight the role of protein disorder in chaperone function and the self-inhibitory role of ordered structures under non-stress conditions, offering new insights for further understanding the protein structure-function paradigm.