Structural Model of a Porphyromonas gingivalis type IX Secretion System Shuttle Complex

Porphyromonas gingivalis is a gram-negative oral anaerobic pathogen and is one of the key causative agents of periodontitis. P. gingivalis utilises a range of virulence factors, including the cysteine protease RgpB, to drive pathogenesis and these are exported and attached to the cell surface via the type IX secretion system (T9SS). All cargo proteins possess a conserved C-terminal signal domain (CTD) which is recognised by the T9SS, and the outer membrane b-barrel protein PorV (PG0027/LptO) can interact with cargo proteins as they are exported to the bacterial surface. Using a combination of solution nuclear magnetic resonance (NMR) spectroscopy, biochemical analyses, machine-learning-based modelling and molecular dynamics (MD) simulations, we present a structural model of a PorV:RgpB-CTD complex from P. gingivalis. This is the first structural insight into CTD recognition by the T9SS and shows how the conserved motifs in the CTD are the primary sites that mediate binding. In PorV, interactions with extracellular surface loops are important for binding the CTD, and together these appear to cradle and lock RgpB-CTD in place. This work provides insight into cargo recognition by PorV but may also have important implications for understanding other aspects of type-IX dependent secretion. (C) 2022 The Author(s). Published by Elsevier Ltd.

Automated summary

This study presents a structural model of the PorV:RgpB-CTD complex from P. gingivalis, providing insight into the recognition mechanism of cargo proteins by the type IX secretion system (T9SS). The study shows that the conserved motifs in the C-terminal signal domain (CTD) are the primary binding sites for T9SS recognition. The interactions of PorV with extracellular surface loops play a crucial role in binding the CTD and securing the RgpB-CTD in place. This research not only advances our understanding of cargo recognition by PorV but also has important implications for other aspects of type-IX dependent secretion.