PDBsum entry 6hv6

Toxin

PDB id: 6hv6

Contents

Protein chain

293 a.a.

Ligands

EDO ×6

EPE

Waters ×184

PDB id:

6hv6

Name:

Toxin

Title:

Crystal structure of patoxp, a cysteine protease-like domain of photorhabdus asymbiotica toxin patox

Structure:

Toxin pau_02230. Chain: a. Synonym: photorhabdus asymbiotica toxin,patox. Engineered: yes. Mutation: yes. Other_details: the catalytic cysteine 1865 was mutated to alanine

Source:

Photorhabdus asymbiotica subsp. Asymbiotica atcc 43949. Organism_taxid: 553480. Gene: pau_02230. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008

Resolution:

2.00Å R-factor: 0.176 R-free: 0.214

Authors:

X.Bogdanovic,C.Wirth,C.Hunte

Key ref:

X.Bogdanovic et al. (2019). A cysteine protease-like domain enhances the cytotoxic effects of the Photorhabdus asymbiotica toxin PaTox. J Biol Chem, 294, 1035-1044. PubMed id: 30478175 DOI: 10.1074/jbc.RA118.005043

Date:

10-Oct-18 Release date: 05-Dec-18

Protein chain

C7BKP9  (PATOX_PHOAA) -  Toxin PAU_02230 from Photorhabdus asymbiotica subsp. asymbiotica (strain ATCC 43949 / 3105-77)

Seq: 2957 a.a.

Struc: 293 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class 1: E.C.2.4.1.- - ?????
• Enzyme class 2: E.C.3.5.1.44 - protein-glutamine glutaminase.
• Reaction: L-glutaminyl-[protein] + H2O = L-glutamyl-[protein] + NH4⁺

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

DOI no: 10.1074/jbc.RA118.005043

J Biol Chem 294:1035-1044 (2019)

PubMed id: 30478175

A cysteine protease-like domain enhances the cytotoxic effects of the Photorhabdus asymbiotica toxin PaTox.

X.Bogdanovic, S.Schneider, N.Levanova, C.Wirth, C.Trillhaase, M.Steinemann, C.Hunte, K.Aktories, T.Jank.

ABSTRACT

The nematode mutualistic bacterium Photorhabdus asymbiotica produces a large virulence-associated multifunctional protein toxin named PaTox. A glycosyltransferase domain and a deamidase domain of this large toxin function as effectors that specifically target host Rho GTPases and heterotrimeric G proteins, respectively. Modification of these intracellular regulators results in toxicity toward insects and mammalian cells. In this study, we identified a cysteine protease-like domain spanning PaTox residues 1844-2114 (PaTox^P), upstream of these two effector domains and characterized by three conserved amino acid residues (Cys-1865, His-1955, and Asp-1975). We determined the crystal structure of the PaTox^P C1865A variant by native single-wavelength anomalous diffraction of sulfur atoms (sulfur-SAD). At 2.0 Å resolution, this structure revealed a catalytic site typical for papain-like cysteine proteases, comprising a catalytic triad, oxyanion hole, and typical secondary structural elements. The PaTox^P structure had highest similarity to that of the AvrPphB protease from Pseudomonas syringae classified as a C58-protease. Furthermore, we observed that PaTox^P shares structural homology also with non-C58-cysteine proteases, deubiquitinases, and deamidases. Upon delivery into insect larvae, PaTox^P alone without full-length PaTox had no toxic effects. Yet, PaTox^P expression in mammalian cells was toxic and enhanced the apoptotic phenotype induced by PaTox in HeLa cells. We propose that PaTox^P is a C58-like cysteine protease module that is essential for full PaTox activity.