CST, 17848S, PTMScan® Propionyl-Lysine [Prop-K] Kit

PTMScan for studying in the research area. Product Usage Information Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan ® Motif Antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan ® IAP Buffer #9993 included in the kit. Storage Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: PTMScan Background Lysine is subject to a wide array of regulatory post-translational modifications due to its positively charged ε-amino group side chain. The most prevalent of these are ubiquitination and acetylation, which are highly conserved among prokaryotes and eukaryotes (1,2). Acyl group transfer from the metabolic intermediates acetyl-, succinyl-, malonyl-, glutaryl-, butyryl-, propionyl-, and crotonyl-CoA all neutralize lysine's positive charge and confer structural alterations affecting substrate protein function. Lysine acetylation is catalyzed by histone acetyltransferases, HATs, using acetyl-CoA as a cofactor (3,4). Deacylation is mediated by histone deacetylases, HDACs 1-11, and NAD-dependent Sirtuins 1-7. Some sirtuins have little to no deacetylase activity, suggesting that they are better suited for other acyl lysine substrates (5). Protein propionyl and butyryl transferase activity has been reported for p300 and CREB-binding protein, two acetyltransferases that can autoacylate as well as target histone proteins and p53 . Sirt1 (Sir2 in yeast) has been shown to have depropionylase activity and may be a major eukaryotic depropionylase (6,7). In the cytosol, acetyl-CoA carboxylase (ACC) converts acetyl-CoA to Malonyl-CoA and the reverse reaction is catalyzed by Malonyl-CoA decarboxylase (MCD), but in the mitochondria, propionyl-CoA carboxylase takes the role of ACC. Both MCD and ACC are regulated by AMPK, glucose levels, and insulin, underscoring their importance in intermediary metabolism (8). Alternate Names Acyl; Acylation; Propionyl; PTMScan