Abstract

The human protease family HtrA is responsible for preventing protein misfolding and mislocalization, and a key player in several cellular processes. Among these, HtrA1 is implicated in several cancers, cerebrovascular disease and age-related macular degeneration. HtrA1 activation, although very relevant for drug-targeting this protease, remains poorly characterized. Our work provides a mechanistic step-by-step description of HtrA1 activation and regulation. We report that the HtrA1 trimer is regulated by an allosteric mechanism by which monomers relay the activation signal to each other, in a PDZ-domain independent fashion. Notably, we show that inhibitor binding is precluded if HtrA1 monomers cannot communicate with each other. Our study establishes how HtrA1 oligomerization plays a fundamental role in proteolytic activity. Moreover, it offers a structural explanation for HtrA1-defective pathologies as well as mechanistic insights into the degradation of complex extracellular fibrils such as tubulin, amyloid beta and tau that belong to the repertoire of HtrA1.\n\nHighlightsO_LIMonomeric HtrA1 is activated by a gating mechanism.\nC_LIO_LITrimeric HtrA1 is regulated by PDZ-independent allosteric monomer cross-talk.\nC_LIO_LIHtrA1 oligomerization is key for proteolytic activity.\nC_LIO_LISubstrate-binding is precluded if monomers cannot communicate with each other.\nC_LI