Identification of antifungal H+-ATPase inhibitors with effect on the plasma membrane potential

Lasse Kjellerup; Sandra Gordon; Karen O'Hanlon Cohrt; William Dalby Brown; Anja Thoe Fuglsang; Anne-Marie Lund Winther

doi:10.1128/AAC.00032-17

Identification of antifungal H⁺-ATPase inhibitors with effect on the plasma membrane potential

Lasse Kjellerup, Sandra Gordon, Karen O'Hanlon Cohrt, William Dalby Brown, Anja Thoe Fuglsang, Anne-Marie Lund Winther

Section for Transport Biology

17 Citations (Scopus)

82 Downloads (Pure)

Abstract

The plasma membrane H⁺-ATPase (Pma1) is an essential fungal protein and a proposed target for new antifungal medications. The compounds in a small-molecule library containing ~191,000 commercially available compounds were screened for their ability to inhibit Saccharomyces cerevisiae plasma membranes containing Pma1. The overall hit rate was 0.2%, corresponding to 407 compounds. These hit compounds were further evaluated for ATPase selectivity and broad-spectrum antifungal activity. Following this work, one Pma1 inhibitor series based on compound 14 and analogs was selected for further evaluation. This compound series was able to depolarize the membrane and inhibit extracellular acidification in intact fungal cells concomitantly with a significant increase in intracellular ATP levels. Collectively, we suggest that these effects may be a common feature of Pma1 inhibitors. Additionally, the work uncovered a dual mechanism for the previously identified cationic peptide BM2, revealing fungal membrane disruption, in addition to Pma1 inhibition. The methods presented here provide a solid platform for the evaluation of Pma1-specific inhibitors in a drug development setting. The present inhibitors could serve as a starting point for the development of new antifungal agents with a novel mode of action.

Original language	English
Article number	e00032-17
Journal	Antimicrobial Agents and Chemotherapy
Volume	61
Issue number	7
Number of pages	14
ISSN	0066-4804
DOIs	https://doi.org/10.1128/AAC.00032-17
Publication status	Published - Jun 2017

Keywords

Journal Article

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Identification of Antifungal H-ATPase Inhibitors with Effect on Plasma Membrane PotentialFinal published version, 1.41 MB

Cite this

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title = "Identification of antifungal H+-ATPase inhibitors with effect on the plasma membrane potential",

abstract = "The plasma membrane H+-ATPase (Pma1) is an essential fungal protein and a proposed target for new antifungal medications. The compounds in a small-molecule library containing ~191,000 commercially available compounds were screened for their ability to inhibit Saccharomyces cerevisiae plasma membranes containing Pma1. The overall hit rate was 0.2%, corresponding to 407 compounds. These hit compounds were further evaluated for ATPase selectivity and broad-spectrum antifungal activity. Following this work, one Pma1 inhibitor series based on compound 14 and analogs was selected for further evaluation. This compound series was able to depolarize the membrane and inhibit extracellular acidification in intact fungal cells concomitantly with a significant increase in intracellular ATP levels. Collectively, we suggest that these effects may be a common feature of Pma1 inhibitors. Additionally, the work uncovered a dual mechanism for the previously identified cationic peptide BM2, revealing fungal membrane disruption, in addition to Pma1 inhibition. The methods presented here provide a solid platform for the evaluation of Pma1-specific inhibitors in a drug development setting. The present inhibitors could serve as a starting point for the development of new antifungal agents with a novel mode of action.",

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author = "Lasse Kjellerup and Sandra Gordon and Cohrt, {Karen O'Hanlon} and Brown, {William Dalby} and Fuglsang, {Anja Thoe} and Winther, {Anne-Marie Lund}",

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year = "2017",

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T1 - Identification of antifungal H+-ATPase inhibitors with effect on the plasma membrane potential

AU - Kjellerup, Lasse

AU - Gordon, Sandra

AU - Cohrt, Karen O'Hanlon

AU - Brown, William Dalby

AU - Fuglsang, Anja Thoe

AU - Winther, Anne-Marie Lund

PY - 2017/6

Y1 - 2017/6

N2 - The plasma membrane H+-ATPase (Pma1) is an essential fungal protein and a proposed target for new antifungal medications. The compounds in a small-molecule library containing ~191,000 commercially available compounds were screened for their ability to inhibit Saccharomyces cerevisiae plasma membranes containing Pma1. The overall hit rate was 0.2%, corresponding to 407 compounds. These hit compounds were further evaluated for ATPase selectivity and broad-spectrum antifungal activity. Following this work, one Pma1 inhibitor series based on compound 14 and analogs was selected for further evaluation. This compound series was able to depolarize the membrane and inhibit extracellular acidification in intact fungal cells concomitantly with a significant increase in intracellular ATP levels. Collectively, we suggest that these effects may be a common feature of Pma1 inhibitors. Additionally, the work uncovered a dual mechanism for the previously identified cationic peptide BM2, revealing fungal membrane disruption, in addition to Pma1 inhibition. The methods presented here provide a solid platform for the evaluation of Pma1-specific inhibitors in a drug development setting. The present inhibitors could serve as a starting point for the development of new antifungal agents with a novel mode of action.

AB - The plasma membrane H+-ATPase (Pma1) is an essential fungal protein and a proposed target for new antifungal medications. The compounds in a small-molecule library containing ~191,000 commercially available compounds were screened for their ability to inhibit Saccharomyces cerevisiae plasma membranes containing Pma1. The overall hit rate was 0.2%, corresponding to 407 compounds. These hit compounds were further evaluated for ATPase selectivity and broad-spectrum antifungal activity. Following this work, one Pma1 inhibitor series based on compound 14 and analogs was selected for further evaluation. This compound series was able to depolarize the membrane and inhibit extracellular acidification in intact fungal cells concomitantly with a significant increase in intracellular ATP levels. Collectively, we suggest that these effects may be a common feature of Pma1 inhibitors. Additionally, the work uncovered a dual mechanism for the previously identified cationic peptide BM2, revealing fungal membrane disruption, in addition to Pma1 inhibition. The methods presented here provide a solid platform for the evaluation of Pma1-specific inhibitors in a drug development setting. The present inhibitors could serve as a starting point for the development of new antifungal agents with a novel mode of action.

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