The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif.
Nontypeable Haemophilus influenzae (NTHi) is a significant pathogen in respiratory disease and otitis media. Important for NTHi survival, colonization and persistence in vivo is the Sap (sensitivity to antimicrobial peptides) ABC transporter system. Current models propose a direct role for Sap in he...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2021-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0256070 |
| _version_ | 1811160736740671488 |
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| author | Petra Lukacik C David Owen Gemma Harris Jani Reddy Bolla Sarah Picaud Irfan Alibay Joanne E Nettleship Louise E Bird Raymond J Owens Philip C Biggin Panagis Filippakopoulos Carol V Robinson Martin A Walsh |
| author_facet | Petra Lukacik C David Owen Gemma Harris Jani Reddy Bolla Sarah Picaud Irfan Alibay Joanne E Nettleship Louise E Bird Raymond J Owens Philip C Biggin Panagis Filippakopoulos Carol V Robinson Martin A Walsh |
| author_sort | Petra Lukacik |
| collection | DOAJ |
| description | Nontypeable Haemophilus influenzae (NTHi) is a significant pathogen in respiratory disease and otitis media. Important for NTHi survival, colonization and persistence in vivo is the Sap (sensitivity to antimicrobial peptides) ABC transporter system. Current models propose a direct role for Sap in heme and antimicrobial peptide (AMP) transport. Here, the crystal structure of SapA, the periplasmic component of Sap, in a closed, ligand bound conformation, is presented. Phylogenetic and cavity volume analysis predicts that the small, hydrophobic SapA central ligand binding cavity is most likely occupied by a hydrophobic di- or tri- peptide. The cavity is of insufficient volume to accommodate heme or folded AMPs. Crystal structures of SapA have identified surface interactions with heme and dsRNA. Heme binds SapA weakly (Kd 282 μM) through a surface exposed histidine, while the dsRNA is coordinated via residues which constitute part of a conserved motif (estimated Kd 4.4 μM). The RNA affinity falls within the range observed for characterized RNA/protein complexes. Overall, we describe in molecular-detail the interactions of SapA with heme and dsRNA and propose a role for SapA in the transport of di- or tri-peptides. |
| first_indexed | 2024-04-10T06:03:39Z |
| format | Article |
| id | doaj.art-3848955c7713409e903d25b26102d894 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-10T06:03:39Z |
| publishDate | 2021-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3848955c7713409e903d25b26102d8942023-03-03T05:31:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032021-01-011610e025607010.1371/journal.pone.0256070The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif.Petra LukacikC David OwenGemma HarrisJani Reddy BollaSarah PicaudIrfan AlibayJoanne E NettleshipLouise E BirdRaymond J OwensPhilip C BigginPanagis FilippakopoulosCarol V RobinsonMartin A WalshNontypeable Haemophilus influenzae (NTHi) is a significant pathogen in respiratory disease and otitis media. Important for NTHi survival, colonization and persistence in vivo is the Sap (sensitivity to antimicrobial peptides) ABC transporter system. Current models propose a direct role for Sap in heme and antimicrobial peptide (AMP) transport. Here, the crystal structure of SapA, the periplasmic component of Sap, in a closed, ligand bound conformation, is presented. Phylogenetic and cavity volume analysis predicts that the small, hydrophobic SapA central ligand binding cavity is most likely occupied by a hydrophobic di- or tri- peptide. The cavity is of insufficient volume to accommodate heme or folded AMPs. Crystal structures of SapA have identified surface interactions with heme and dsRNA. Heme binds SapA weakly (Kd 282 μM) through a surface exposed histidine, while the dsRNA is coordinated via residues which constitute part of a conserved motif (estimated Kd 4.4 μM). The RNA affinity falls within the range observed for characterized RNA/protein complexes. Overall, we describe in molecular-detail the interactions of SapA with heme and dsRNA and propose a role for SapA in the transport of di- or tri-peptides.https://doi.org/10.1371/journal.pone.0256070 |
| spellingShingle | Petra Lukacik C David Owen Gemma Harris Jani Reddy Bolla Sarah Picaud Irfan Alibay Joanne E Nettleship Louise E Bird Raymond J Owens Philip C Biggin Panagis Filippakopoulos Carol V Robinson Martin A Walsh The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif. PLoS ONE |
| title | The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif. |
| title_full | The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif. |
| title_fullStr | The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif. |
| title_full_unstemmed | The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif. |
| title_short | The structure of nontypeable Haemophilus influenzae SapA in a closed conformation reveals a constricted ligand-binding cavity and a novel RNA binding motif. |
| title_sort | structure of nontypeable haemophilus influenzae sapa in a closed conformation reveals a constricted ligand binding cavity and a novel rna binding motif |
| url | https://doi.org/10.1371/journal.pone.0256070 |
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