A rapid and sensitive assay for histone acetyl-transferase activity.
Histone acetyl-transferases (HATs) seem to be key elements in the regulation of transcription. We have designed an enzymatic assay to quantify HAT enzymatic activity. In this assay, the substrate is a peptide corresponding to the 24 first amino acids of histone H4 which is coupled to biotin. After acetylation using [14C]acetyl-CoA, the peptide is purified on streptavidin beads and the associated radioactivity is measured. This assay is sensitive, rapid and convenient.
Ait-Si-Ali
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Laboratoire Oncogénèse, Différenciation et Transduction du Signal, CNRS UPR 9079, IFC-01, 7 rue Guy Moquet, 94801 Villejuif, France.
Ramirez
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Robin
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Trouche
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Harel-Bellan
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D016475:3T3 Cells; D000105:Acetyl Coenzyme A; D000107:Acetylation; D000123:Acetyltransferases / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin; D002250:Carbon Radioisotopes; D051548:Histone Acetyltransferases; D006657:Histones / Q000737:chemistry / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D012680:Sensitivity and Specificity; D019809:Streptavidin; D013379:Substrate Specificity
D002250:Carbon Radioisotopes; D006657:Histones; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D001710:Biotin; D000105:Acetyl Coenzyme A; D019809:Streptavidin; D000123:Acetyltransferases; D051548:Histone Acetyltransferases