title: A structure-based model of substrate discrimination by a noncanonical PDZ tandem in the intramembrane-cleaving protease RseP.
abstract: During the extracytoplasmic stress response in Escherichia coli, the intramembrane protease RseP cleaves the anti-σ(E) protein RseA only after the membrane-anchored protease DegS truncates the periplasmic part of RseA that suppresses the action of RseP. Here we analyzed the three-dimensional structure of the two tandemly arranged PSD-95/Dlg/ZO-1 (PDZ) domains (PDZ tandem) present in the periplasmic region of RseP and revealed that the two putative ligand-binding grooves constitute a single pocket-like structure that would lie just above the active center sequestrated within the membrane. Complete removal of the PDZ tandem from RseP led to the intramembrane cleavage of RseA without prior truncation by DegS. Furthermore, mutations expected to destabilize the tertiary structure of the PDZ tandem also caused the deregulation of the sequential cleavage. These observations suggest that the PDZ tandem serves as a size-exclusion filter to accommodate the truncated form of RseA into the active center.
authors.affiliation: Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Hizukuri
authors.affiliation: Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Oda
authors.affiliation: Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Sanae
authors.initials: S
authors.lastname: Tabata
authors.affiliation: Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Tamura-Kawakami
authors.affiliation: Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Rika
authors.initials: R
authors.lastname: Oi
authors.affiliation: Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama 230-0045, Japan; RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Sato
authors.affiliation: Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Junichi
authors.initials: J
authors.lastname: Takagi
authors.affiliation: Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Akiyama
authors.affiliation: Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama 230-0045, Japan. Electronic address: nogi@tsurumi.yokohama-cu.ac.jp.
authors.forename: Terukazu
authors.initials: T
authors.lastname: Nogi
mesh_terms: D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D010450:Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D008024:Ligands; D008565:Membrane Proteins / Q000378:metabolism*; D009154:Mutation; D011485:Protein Binding
chemical_list: D029968:Escherichia coli Proteins; D008024:Ligands; D008565:Membrane Proteins; D010450:Endopeptidases; C440001:RseP protein, E coli