X-ray and Cryo-electron Microscopy Structures of Monalysin Pore-forming Toxin Reveal Multimerization of the Pro-form.

Details

Serval ID
serval:BIB_1426D6EC1F2B
Type
Article: article from journal or magazin.
Collection
Publications
Title
X-ray and Cryo-electron Microscopy Structures of Monalysin Pore-forming Toxin Reveal Multimerization of the Pro-form.
Journal
Journal of Biological Chemistry
Author(s)
Leone P., Bebeacua C., Opota O., Kellenberger C., Klaholz B., Orlov I., Cambillau C., Lemaitre B., Roussel A.
ISSN
1083-351X (Electronic)
ISSN-L
0021-9258
Publication state
Published
Issued date
2015
Volume
290
Number
21
Pages
13191-13201
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
β-Barrel pore-forming toxins (β-PFT), a large family of bacterial toxins, are generally secreted as water-soluble monomers and can form oligomeric pores in membranes following proteolytic cleavage and interaction with cell surface receptors. Monalysin has been recently identified as a β-PFT that contributes to the virulence of Pseudomonas entomophila against Drosophila. It is secreted as a pro-protein that becomes active upon cleavage. Here we report the crystal and cryo-electron microscopy structure of the pro-form of Monalysin as well as the crystal structures of the cleaved form and of an inactive mutant lacking the membrane-spanning region. The overall structure of Monalysin displays an elongated shape, which resembles those of β-pore-forming toxins, such as Aerolysin, but is devoid of a receptor-binding domain. X-ray crystallography, cryo-electron microscopy, and light-scattering studies show that pro-Monalysin forms a stable doughnut-like 18-mer complex composed of two disk-shaped nonamers held together by N-terminal swapping of the pro-peptides. This observation is in contrast with the monomeric pro-form of the other β-PFTs that are receptor-dependent for membrane interaction. The membrane-spanning region of pro-Monalysin is fully buried in the center of the doughnut, suggesting that upon cleavage of pro-peptides, the two disk-shaped nonamers can, and have to, dissociate to leave the transmembrane segments free to deploy and lead to pore formation. In contrast with other toxins, the delivery of 18 subunits at once, nearby the cell surface, may be used to bypass the requirement of receptor-dependent concentration to reach the threshold for oligomerization into the pore-forming complex.
Keywords
Amino Acid Sequence, Bacterial Toxins/chemistry, Bacterial Toxins/genetics, Cell Membrane, Cryoelectron Microscopy/methods, Crystallization, Crystallography, X-Ray, Models, Molecular, Molecular Sequence Data, Mutation/genetics, Pore Forming Cytotoxic Proteins/chemistry, Pore Forming Cytotoxic Proteins/genetics, Protein Binding, Protein Multimerization, Protein Structure, Tertiary, Pseudomonas/genetics, Pseudomonas/growth & development, Sequence Homology, Amino Acid
Pubmed
Open Access
Yes
Create date
02/11/2015 17:19
Last modification date
20/08/2019 12:42
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