N-myristoyltransferase is a cell wall target in Aspergillus fumigatus

Wenxia Fang; Olawale G Raimi; David E Blair; Justin R Harrison; Deborah E A Lockhart; Leah S Torrie; Gian Filippo Ruda; Paul G Wyatt; Ian H Gilbert; Daan M F van Aalten

doi:10.1021/cb5008647

N-myristoyltransferase is a cell wall target in Aspergillus fumigatus

Wenxia Fang, Olawale G Raimi, David E Blair, Justin R Harrison, Deborah E A Lockhart, Leah S Torrie, Gian Filippo Ruda, Paul G Wyatt, Ian H Gilbert, Daan M F van Aalten

University of Dundee

Research output: Contribution to journal › Article › peer-review

36 Citations (Scopus)

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Abstract

Treatment of filamentous fungal infections relies on a limited repertoire of antifungal agents. Compounds possessing novel modes of action are urgently required. N-myristoylation is a ubiquitous modification of eukaryotic proteins. The enzyme N-myristoyltransferase (NMT) has been considered a potential therapeutic target in protozoa and yeasts. Here, we show that the filamentous fungal pathogen Aspergillus fumigatus possesses an active NMT enzyme that is essential for survival. Surprisingly, partial repression of the gene revealed downstream effects of N-myristoylation on cell wall morphology. Screening a library of inhibitors led to the discovery of a pyrazole sulphonamide compound that inhibits the enzyme and is fungicidal under partially repressive nmt conditions. Together with a crystallographic complex showing the inhibitor binding in the peptide substrate pocket, we provide evidence of NMT being a potential drug target in A. fumigatus.

Original language	English
Pages (from-to)	1425-1434
Number of pages	10
Journal	ACS Chemical Biology
Volume	10
Issue number	6
Early online date	27 Feb 2015
DOIs	https://doi.org/10.1021/cb5008647
Publication status	Published - 19 Jun 2015
Externally published	Yes

Bibliographical note

The authors would like to thank the European Synchrotron Radiation facility (ESRF), Grenoble, for beamline time. We thank J. James of the College of Life Sciences Microscopy Facility for his help with electron microscopy. We also thank F. Eisenhaber, Bioinformatics Institute Singapore, for predicting myristoylated proteins. This work was funded by an MRC Programme Grant (M004139) and a Wellcome Trust Senior Research Fellowship (WT087590MA) to D.M.F.v.A. D.E.A.L was supported by an MRC Clinical Research Training Fellowship (G1100430). We also would like to acknowledge the Wellcome Trust (WT077705, WT083481) for support.

Supporting Figures S1−S3, Supporting Tables S1−S4, and further details of experimental procedures. This material is available free of charge via the Internet from http://pubs.acs.org.

Keywords

Acyltransferases
Aminopyridines
Antifungal Agents
Aspergillus fumigatus
Catalytic Domain
Cell Wall
Crystallography, X-Ray
Fungal Proteins
Kinetics
Microbial Sensitivity Tests
Protein Binding
Protein Processing, Post-Translational
Protein Structure, Secondary
Pyrazoles
Structure-Activity Relationship
Sulfonamides
Journal Article
Research Support, Non-U.S. Gov't

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fang_etal_ACSCB_N_Myristoyltransferase_Is_VOR
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Cite this

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title = "N-myristoyltransferase is a cell wall target in Aspergillus fumigatus",

abstract = "Treatment of filamentous fungal infections relies on a limited repertoire of antifungal agents. Compounds possessing novel modes of action are urgently required. N-myristoylation is a ubiquitous modification of eukaryotic proteins. The enzyme N-myristoyltransferase (NMT) has been considered a potential therapeutic target in protozoa and yeasts. Here, we show that the filamentous fungal pathogen Aspergillus fumigatus possesses an active NMT enzyme that is essential for survival. Surprisingly, partial repression of the gene revealed downstream effects of N-myristoylation on cell wall morphology. Screening a library of inhibitors led to the discovery of a pyrazole sulphonamide compound that inhibits the enzyme and is fungicidal under partially repressive nmt conditions. Together with a crystallographic complex showing the inhibitor binding in the peptide substrate pocket, we provide evidence of NMT being a potential drug target in A. fumigatus.",

keywords = "Acyltransferases, Aminopyridines, Antifungal Agents, Aspergillus fumigatus, Catalytic Domain, Cell Wall, Crystallography, X-Ray, Fungal Proteins, Kinetics, Microbial Sensitivity Tests, Protein Binding, Protein Processing, Post-Translational, Protein Structure, Secondary, Pyrazoles, Structure-Activity Relationship, Sulfonamides, Journal Article, Research Support, Non-U.S. Gov't",

author = "Wenxia Fang and Raimi, {Olawale G} and Blair, {David E} and Harrison, {Justin R} and Lockhart, {Deborah E A} and Torrie, {Leah S} and Ruda, {Gian Filippo} and Wyatt, {Paul G} and Gilbert, {Ian H} and {van Aalten}, {Daan M F}",

note = "The authors would like to thank the European Synchrotron Radiation facility (ESRF), Grenoble, for beamline time. We thank J. James of the College of Life Sciences Microscopy Facility for his help with electron microscopy. We also thank F. Eisenhaber, Bioinformatics Institute Singapore, for predicting myristoylated proteins. This work was funded by an MRC Programme Grant (M004139) and a Wellcome Trust Senior Research Fellowship (WT087590MA) to D.M.F.v.A. D.E.A.L was supported by an MRC Clinical Research Training Fellowship (G1100430). We also would like to acknowledge the Wellcome Trust (WT077705, WT083481) for support. Supporting Figures S1−S3, Supporting Tables S1−S4, and further details of experimental procedures. This material is available free of charge via the Internet from http://pubs.acs.org.",

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doi = "10.1021/cb5008647",

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AU - Fang, Wenxia

AU - Raimi, Olawale G

AU - Blair, David E

AU - Harrison, Justin R

AU - Lockhart, Deborah E A

AU - Torrie, Leah S

AU - Ruda, Gian Filippo

AU - Wyatt, Paul G

AU - Gilbert, Ian H

AU - van Aalten, Daan M F

N1 - The authors would like to thank the European Synchrotron Radiation facility (ESRF), Grenoble, for beamline time. We thank J. James of the College of Life Sciences Microscopy Facility for his help with electron microscopy. We also thank F. Eisenhaber, Bioinformatics Institute Singapore, for predicting myristoylated proteins. This work was funded by an MRC Programme Grant (M004139) and a Wellcome Trust Senior Research Fellowship (WT087590MA) to D.M.F.v.A. D.E.A.L was supported by an MRC Clinical Research Training Fellowship (G1100430). We also would like to acknowledge the Wellcome Trust (WT077705, WT083481) for support. Supporting Figures S1−S3, Supporting Tables S1−S4, and further details of experimental procedures. This material is available free of charge via the Internet from http://pubs.acs.org.

PY - 2015/6/19

Y1 - 2015/6/19

N2 - Treatment of filamentous fungal infections relies on a limited repertoire of antifungal agents. Compounds possessing novel modes of action are urgently required. N-myristoylation is a ubiquitous modification of eukaryotic proteins. The enzyme N-myristoyltransferase (NMT) has been considered a potential therapeutic target in protozoa and yeasts. Here, we show that the filamentous fungal pathogen Aspergillus fumigatus possesses an active NMT enzyme that is essential for survival. Surprisingly, partial repression of the gene revealed downstream effects of N-myristoylation on cell wall morphology. Screening a library of inhibitors led to the discovery of a pyrazole sulphonamide compound that inhibits the enzyme and is fungicidal under partially repressive nmt conditions. Together with a crystallographic complex showing the inhibitor binding in the peptide substrate pocket, we provide evidence of NMT being a potential drug target in A. fumigatus.

AB - Treatment of filamentous fungal infections relies on a limited repertoire of antifungal agents. Compounds possessing novel modes of action are urgently required. N-myristoylation is a ubiquitous modification of eukaryotic proteins. The enzyme N-myristoyltransferase (NMT) has been considered a potential therapeutic target in protozoa and yeasts. Here, we show that the filamentous fungal pathogen Aspergillus fumigatus possesses an active NMT enzyme that is essential for survival. Surprisingly, partial repression of the gene revealed downstream effects of N-myristoylation on cell wall morphology. Screening a library of inhibitors led to the discovery of a pyrazole sulphonamide compound that inhibits the enzyme and is fungicidal under partially repressive nmt conditions. Together with a crystallographic complex showing the inhibitor binding in the peptide substrate pocket, we provide evidence of NMT being a potential drug target in A. fumigatus.

KW - Acyltransferases

KW - Aminopyridines

KW - Antifungal Agents

KW - Aspergillus fumigatus

KW - Catalytic Domain

KW - Cell Wall

KW - Crystallography, X-Ray

KW - Fungal Proteins

KW - Kinetics

KW - Microbial Sensitivity Tests

KW - Protein Binding

KW - Protein Processing, Post-Translational

KW - Protein Structure, Secondary

KW - Pyrazoles

KW - Structure-Activity Relationship

KW - Sulfonamides

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1021/cb5008647

DO - 10.1021/cb5008647

M3 - Article

C2 - 25706802

SN - 1554-8929

VL - 10

SP - 1425

EP - 1434

JO - ACS Chemical Biology

JF - ACS Chemical Biology

IS - 6

ER -

N-myristoyltransferase is a cell wall target in Aspergillus fumigatus

Abstract

Bibliographical note

Keywords

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