High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers.
Pore-forming proteins (PFPs) represent a functionally important protein family, that are found in organisms from viruses to humans. As a major branch of PFPs, bacteria pore-forming toxins (PFTs) permeabilize membranes and usually cause the death of target cells. E. coli hemolysin ClyA is the first m...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2019-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0213423 |
| _version_ | 1797209651720224768 |
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| author | Wei Peng Marcela de Souza Santos Yang Li Diana R Tomchick Kim Orth |
| author_facet | Wei Peng Marcela de Souza Santos Yang Li Diana R Tomchick Kim Orth |
| author_sort | Wei Peng |
| collection | DOAJ |
| description | Pore-forming proteins (PFPs) represent a functionally important protein family, that are found in organisms from viruses to humans. As a major branch of PFPs, bacteria pore-forming toxins (PFTs) permeabilize membranes and usually cause the death of target cells. E. coli hemolysin ClyA is the first member with the pore complex structure solved among α-PFTs, employing α-helices as transmembrane elements. ClyA is proposed to form pores composed of various numbers of protomers. With high-resolution cryo-EM structures, we observe that ClyA pore complexes can exist as newly confirmed oligomers of a tridecamer and a tetradecamer, at estimated resolutions of 3.2 Å and 4.3 Å, respectively. The 2.8 Å cryo-EM structure of a dodecamer dramatically improves the existing structural model. Structural analysis indicates that protomers from distinct oligomers resemble each other and neighboring protomers adopt a conserved interaction mode. We also show a stabilized intermediate state of ClyA during the transition process from soluble monomers to pore complexes. Unexpectedly, even without the formation of mature pore complexes, ClyA can permeabilize membranes and allow leakage of particles less than ~400 Daltons. In addition, we are the first to show that ClyA forms pore complexes in the presence of cholesterol within artificial liposomes. These findings provide new mechanistic insights into the dynamic process of pore assembly for the prototypical α-PFT ClyA. |
| first_indexed | 2024-04-24T09:58:06Z |
| format | Article |
| id | doaj.art-d6ac5573073d4aa69b01bd4ad0df7052 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-24T09:58:06Z |
| publishDate | 2019-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-d6ac5573073d4aa69b01bd4ad0df70522024-04-14T05:31:57ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-01145e021342310.1371/journal.pone.0213423High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers.Wei PengMarcela de Souza SantosYang LiDiana R TomchickKim OrthPore-forming proteins (PFPs) represent a functionally important protein family, that are found in organisms from viruses to humans. As a major branch of PFPs, bacteria pore-forming toxins (PFTs) permeabilize membranes and usually cause the death of target cells. E. coli hemolysin ClyA is the first member with the pore complex structure solved among α-PFTs, employing α-helices as transmembrane elements. ClyA is proposed to form pores composed of various numbers of protomers. With high-resolution cryo-EM structures, we observe that ClyA pore complexes can exist as newly confirmed oligomers of a tridecamer and a tetradecamer, at estimated resolutions of 3.2 Å and 4.3 Å, respectively. The 2.8 Å cryo-EM structure of a dodecamer dramatically improves the existing structural model. Structural analysis indicates that protomers from distinct oligomers resemble each other and neighboring protomers adopt a conserved interaction mode. We also show a stabilized intermediate state of ClyA during the transition process from soluble monomers to pore complexes. Unexpectedly, even without the formation of mature pore complexes, ClyA can permeabilize membranes and allow leakage of particles less than ~400 Daltons. In addition, we are the first to show that ClyA forms pore complexes in the presence of cholesterol within artificial liposomes. These findings provide new mechanistic insights into the dynamic process of pore assembly for the prototypical α-PFT ClyA.https://doi.org/10.1371/journal.pone.0213423 |
| spellingShingle | Wei Peng Marcela de Souza Santos Yang Li Diana R Tomchick Kim Orth High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. PLoS ONE |
| title | High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. |
| title_full | High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. |
| title_fullStr | High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. |
| title_full_unstemmed | High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. |
| title_short | High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. |
| title_sort | high resolution cryo em structures of the e coli hemolysin clya oligomers |
| url | https://doi.org/10.1371/journal.pone.0213423 |
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