Activity-based substrate profiling for Gcn5-related N-acetyltransferases: the use of chloroacetyl-coenzyme A to identify protein substrates.
The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10 000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His8) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the alpha-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes.
Yu
Michael
M
Department of Biochemistry, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
de Carvalho
Luiz Pedro Sorio
LP
Sun
Guangxing
G
Blanchard
John S
JS
D000105:Acetyl Coenzyme A / Q000737:chemistry*; D000107:Acetylation; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D051548:Histone Acetyltransferases / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D019779:Salmonella enterica / Q000201:enzymology / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; C026772:chloroacetyl coenzyme A; D000105:Acetyl Coenzyme A; D051548:Histone Acetyltransferases