9J82

Cryo-EM structure of wild type Aquifex aeolicus RseP in complex with Fab

Summary for 9J82

• Entry DOI: 10.2210/pdb9j82/pdb
• EMDB information: 61213
• Descriptor: Putative zinc metalloprotease aq_1964, VH-CH1 region of mouse monoclonal antibody IgG 4A9, L chain of mouse monoclonal antibody IgG 4A9, ... (4 entities in total)
• Functional Keywords: protease, hydrolase
• Biological source: Aquifex aeolicus VF5; More
• Total number of polymer chains: 3
• Total formula weight: 98686.33

Authors

Asahi, K.,Hirose, M.,Aruga, R.,Kato, T.,Nogi, T. (deposition date: 2024-08-20, release date: 2025-03-05, Last modification date: 2025-03-19)

Primary citation

Asahi, K.,Hirose, M.,Aruga, R.,Shimizu, Y.,Tajiri, M.,Tanaka, T.,Adachi, Y.,Tanaka, Y.,Kaneko, M.K.,Kato, Y.,Akashi, S.,Akiyama, Y.,Hizukuri, Y.,Kato, T.,Nogi, T.
Cryo-EM structure of the bacterial intramembrane metalloprotease RseP in the substrate-bound state.
Sci Adv, 11:eadu0925-eadu0925, 2025

PubMed Abstract: Site-2 proteases (S2Ps), conserved intramembrane metalloproteases that maintain cellular homeostasis, are associated with chronic infection and persistence leading to multidrug resistance in bacterial pathogens. A structural model of how S2Ps discriminate and accommodate substrates could help us develop selective antimicrobial agents. We previously proposed that the S2P RseP unwinds helical substrate segments before cleavage, but the mechanism for accommodating a full-length membrane-spanning substrate remained unclear. Our present cryo-EM analysis of RseP (RseP) revealed that a substrate-like membrane protein fragment from the expression host occupied the active site while spanning a transmembrane cavity that is inaccessible via lateral diffusion. Furthermore, in vivo photocrosslinking supported that this substrate accommodation mode is recapitulated on the cell membrane. Our results suggest that the substrate accommodation by threading through a conserved membrane-associated region stabilizes the substrate-complex and contributes to substrate discrimination on the membrane.

PubMed: 40009668
DOI: 10.1126/sciadv.adu0925

Experimental method

ELECTRON MICROSCOPY (3.95 Å)