Journal
/
Biochemistry
/
Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay
JoVE Journal
Biochemistry
This content is Free Access.
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

DOI:
10.3791/59527-v

•

09:07 min

•

May 01, 2019

•

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

Chapters

  • 00:04Title
  • 00:53Inoculum Preparation
  • 01:34Growth and Harvesting of Mycelia
  • 02:36Preparation of the Protein Extract and Equilibration of the IgG Resin
  • 04:40Batch Purification of TAP-tagged HDAC
  • 06:50Results: Analysis of the Purification and Enzymatic Activity Assay
  • 08:29Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/59527?article_type=v

A Fluorescence-based Assay of Phospholipid Scramblase Activity
check_url/59527?article_type=v

Analysis of Histone Antibody Specificity with Peptide Microarrays
check_url/59527?article_type=v

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity
check_url/59527?article_type=v

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
check_url/59527?article_type=v

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
check_url/59527?article_type=v

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
check_url/59527?article_type=v

A Colorimetric Assay of Citrate Synthase Activity in Drosophila Melanogaster
check_url/59527?article_type=v

A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
check_url/59527?article_type=v

Isolation of Histone from Sorghum Leaf Tissue for Top Down Mass Spectrometry Profiling of Potential Epigenetic Markers
check_url/59527?article_type=v

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1

Read Article