Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane

Daptomycin is a potent lipopeptide antibiotic used in the treatment of life-threatening Gram-positive infections, but the molecular mechanism of its interaction with bacterial membrane remains unclear. Here, we show that this interaction is divided into two stages, of which the first is a fast and r...

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Main Authors: Pragyansree Machhua, Vignesh Gopalakrishnan Unnithan, Yu Liu, Yiping Jiang, Lingfeng Zhang, Zhihong Guo
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2025-06-01
Series:eLife
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Online Access:https://elifesciences.org/articles/93267
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author Pragyansree Machhua
Vignesh Gopalakrishnan Unnithan
Yu Liu
Yiping Jiang
Lingfeng Zhang
Zhihong Guo
author_facet Pragyansree Machhua
Vignesh Gopalakrishnan Unnithan
Yu Liu
Yiping Jiang
Lingfeng Zhang
Zhihong Guo
author_sort Pragyansree Machhua
collection DOAJ
description Daptomycin is a potent lipopeptide antibiotic used in the treatment of life-threatening Gram-positive infections, but the molecular mechanism of its interaction with bacterial membrane remains unclear. Here, we show that this interaction is divided into two stages, of which the first is a fast and reversible binding of the drug to phospholipid membrane in milliseconds, and the second is a slow and irreversible insertion into membrane in minutes, only in the presence of the bacteria-specific lipid phosphatidylglycerol, to a saturating point where the ratio of the drug to phosphatidylglycerol is 1:2. Fluorescence-based titration showed that the antibiotic simultaneously binds two molecules of phosphatidylglycerol with a nanomolar binding affinity in the presence of calcium ion. The resulting stable complex is easily formed in a test tube and readily isolated from the membrane of drug-treated bacterial cells, strongly supporting a unique drug uptake mechanism in which daptomycin forms a stable multicomponent complex with calcium and phosphatidylglycerol. Revelation of this novel uptake mechanism provides fresh insights into the mode of action of daptomycin and paves the way to new strategies to attenuate resistance to the drug.
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institution OA Journals
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publisher eLife Sciences Publications Ltd
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series eLife
spelling doaj-art-73185282d45643a7bf5ff189d220d7182025-08-20T02:09:08ZengeLife Sciences Publications LtdeLife2050-084X2025-06-011310.7554/eLife.93267Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membranePragyansree Machhua0https://orcid.org/0000-0001-9425-130XVignesh Gopalakrishnan Unnithan1Yu Liu2https://orcid.org/0000-0001-6179-7699Yiping Jiang3Lingfeng Zhang4https://orcid.org/0009-0009-5282-8022Zhihong Guo5https://orcid.org/0000-0003-0374-8412Shenzhen Research Institute and Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, ChinaShenzhen Research Institute and Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, ChinaShenzhen Research Institute and Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, ChinaShenzhen Research Institute and Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, ChinaShenzhen Research Institute and Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, ChinaShenzhen Research Institute and Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, ChinaDaptomycin is a potent lipopeptide antibiotic used in the treatment of life-threatening Gram-positive infections, but the molecular mechanism of its interaction with bacterial membrane remains unclear. Here, we show that this interaction is divided into two stages, of which the first is a fast and reversible binding of the drug to phospholipid membrane in milliseconds, and the second is a slow and irreversible insertion into membrane in minutes, only in the presence of the bacteria-specific lipid phosphatidylglycerol, to a saturating point where the ratio of the drug to phosphatidylglycerol is 1:2. Fluorescence-based titration showed that the antibiotic simultaneously binds two molecules of phosphatidylglycerol with a nanomolar binding affinity in the presence of calcium ion. The resulting stable complex is easily formed in a test tube and readily isolated from the membrane of drug-treated bacterial cells, strongly supporting a unique drug uptake mechanism in which daptomycin forms a stable multicomponent complex with calcium and phosphatidylglycerol. Revelation of this novel uptake mechanism provides fresh insights into the mode of action of daptomycin and paves the way to new strategies to attenuate resistance to the drug.https://elifesciences.org/articles/93267daptomycinmode of actionresistancephospholipidsmembrane insertion
spellingShingle Pragyansree Machhua
Vignesh Gopalakrishnan Unnithan
Yu Liu
Yiping Jiang
Lingfeng Zhang
Zhihong Guo
Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
eLife
daptomycin
mode of action
resistance
phospholipids
membrane insertion
title Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
title_full Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
title_fullStr Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
title_full_unstemmed Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
title_short Daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
title_sort daptomycin forms a stable complex with phosphatidylglycerol for selective uptake to bacterial membrane
topic daptomycin
mode of action
resistance
phospholipids
membrane insertion
url https://elifesciences.org/articles/93267
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AT yuliu daptomycinformsastablecomplexwithphosphatidylglycerolforselectiveuptaketobacterialmembrane
AT yipingjiang daptomycinformsastablecomplexwithphosphatidylglycerolforselectiveuptaketobacterialmembrane
AT lingfengzhang daptomycinformsastablecomplexwithphosphatidylglycerolforselectiveuptaketobacterialmembrane
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