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Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay

Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay

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6,523 Views

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09:07 min

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May 01, 2019

DOI:

10.3791/59527-v

DOI:
10.3791/59527-v

•

09:07 min
May 01, 2019

•

2 Views
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1Division of Molecular Biology, Biocenter,Medical University of Innsbruck, 2Biology Department,Maynooth University, 3Maynooth University Human Health Research Institute

Summary

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Class 1 histone deacetylases (HDACs) like RpdA have gained importance as potential targets to treat fungal infections. Here we present a protocol for the specific enrichment of TAP-tagged RpdA combined with an HDAC activity assay that allows in vitro efficacy testing of histone deacetylase inhibitors.

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TAP-taggedHistone DeacetylaseAspergillus NidulansEnzymatic Activity AssayClass 1 Histone DeacetylasesRpdAFungal InfectionsHDAC ComplexesFungal-specific Deacetylase InhibitorsConidiaMyceliaLyophilizationBall MillExtraction Buffer

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Bauer, I., Pidroni, A., Bayram, Ö., Brosch, G., Graessle, S. Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay. J. Vis. Exp. (147), e59527, doi:10.3791/59527 (2019).

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