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.
Laboratoire Oncogénèse, Différenciation et Transduction du Signal, CNRS UPR 9079, IFC-01, 7 rue Guy Moquet, 94801 Villejuif, France.
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S
Ait-Si-Ali
S
S
Ramirez
P
P
Robin
D
D
Trouche
A
A
Harel-Bellan
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