Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae
Abstract Heme internalization by pathogenic bacteria inside a human host to accomplish the requirement of iron for important cellular processes is of paramount importance. Despite this, the mechanism of heme import by the ATP-binding-cassette (ABC) transporter HutCD in Vibrio cholerae remains unexpl...
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| Format: | Article |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-11213-9 |
| _version_ | 1811306284184502272 |
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| author | Indrila Saha Shrestha Chakraborty Shubhangi Agarwal Peeali Mukherjee Biplab Ghosh Jhimli Dasgupta |
| author_facet | Indrila Saha Shrestha Chakraborty Shubhangi Agarwal Peeali Mukherjee Biplab Ghosh Jhimli Dasgupta |
| author_sort | Indrila Saha |
| collection | DOAJ |
| description | Abstract Heme internalization by pathogenic bacteria inside a human host to accomplish the requirement of iron for important cellular processes is of paramount importance. Despite this, the mechanism of heme import by the ATP-binding-cassette (ABC) transporter HutCD in Vibrio cholerae remains unexplored. We have performed biochemical studies on ATPase HutD and its mutants, along with molecular modelling, docking and unbiased all-atom MD simulations on lipid-solvated models of permease-ATPase complex HutCD. The results demonstrated mechanisms of ATP binding/hydrolysis and trapped transient and global conformational changes in HutCD, necessary for heme internalization. ATPase HutD forms a dimer, independent of the permease HutC. Each HutD monomer canonically binds ATP in a 1:1 stoichiometry. MD simulations demonstrated that a rotational motion of HutC dimer occurs synchronously with the inter-dimeric D-loop interactions of HutDs. F151 of TM4–TM5 loop of HutC, packs with ATP and Y15 of HutD, initiating ‘cytoplasmic gate opening’ which mimics an ‘outward-facing’ to ‘inward-facing’ conformational switching upon ATP hydrolysis. The simulation on ‘inward-facing’ HutCD culminates to an ‘occluded’ state. The simulation on heme-docked HutCD indicated that the event of heme release occurs in ATP-free ‘inward-facing’ state. Gradual conformational changes of the TM5 helices of HutC towards the ‘occluded’ state facilitate ejection of heme. |
| first_indexed | 2024-04-13T08:41:35Z |
| format | Article |
| id | doaj.art-e896587dee514b679cbf414135685a83 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-13T08:41:35Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-e896587dee514b679cbf414135685a832022-12-22T02:53:53ZengNature PortfolioScientific Reports2045-23222022-05-0112111810.1038/s41598-022-11213-9Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio choleraeIndrila Saha0Shrestha Chakraborty1Shubhangi Agarwal2Peeali Mukherjee3Biplab Ghosh4Jhimli Dasgupta5Department of Biotechnology, St. Xavier’s College (Autonomous)Department of Biotechnology, St. Xavier’s College (Autonomous)Department of Biotechnology, St. Xavier’s College (Autonomous)Department of Biotechnology, St. Xavier’s College (Autonomous)Macromolecular Crystallography Section, Beamline Development & Application Section, Bhabha Atomic Research CenterDepartment of Biotechnology, St. Xavier’s College (Autonomous)Abstract Heme internalization by pathogenic bacteria inside a human host to accomplish the requirement of iron for important cellular processes is of paramount importance. Despite this, the mechanism of heme import by the ATP-binding-cassette (ABC) transporter HutCD in Vibrio cholerae remains unexplored. We have performed biochemical studies on ATPase HutD and its mutants, along with molecular modelling, docking and unbiased all-atom MD simulations on lipid-solvated models of permease-ATPase complex HutCD. The results demonstrated mechanisms of ATP binding/hydrolysis and trapped transient and global conformational changes in HutCD, necessary for heme internalization. ATPase HutD forms a dimer, independent of the permease HutC. Each HutD monomer canonically binds ATP in a 1:1 stoichiometry. MD simulations demonstrated that a rotational motion of HutC dimer occurs synchronously with the inter-dimeric D-loop interactions of HutDs. F151 of TM4–TM5 loop of HutC, packs with ATP and Y15 of HutD, initiating ‘cytoplasmic gate opening’ which mimics an ‘outward-facing’ to ‘inward-facing’ conformational switching upon ATP hydrolysis. The simulation on ‘inward-facing’ HutCD culminates to an ‘occluded’ state. The simulation on heme-docked HutCD indicated that the event of heme release occurs in ATP-free ‘inward-facing’ state. Gradual conformational changes of the TM5 helices of HutC towards the ‘occluded’ state facilitate ejection of heme.https://doi.org/10.1038/s41598-022-11213-9 |
| spellingShingle | Indrila Saha Shrestha Chakraborty Shubhangi Agarwal Peeali Mukherjee Biplab Ghosh Jhimli Dasgupta Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae Scientific Reports |
| title | Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae |
| title_full | Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae |
| title_fullStr | Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae |
| title_full_unstemmed | Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae |
| title_short | Mechanistic insights of ABC importer HutCD involved in heme internalization by Vibrio cholerae |
| title_sort | mechanistic insights of abc importer hutcd involved in heme internalization by vibrio cholerae |
| url | https://doi.org/10.1038/s41598-022-11213-9 |
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