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XML generation date: 2024-09-04 22:38:12.165
Product last modified at: 2024-09-05T08:00:17.301Z
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PDP - Template Name: PTMScan (with Pricing)
PDP - Template ID: *******57cbce3

PTMScan® Glutaryl-Lysine [Glut-K] Kit #26101

Additional Information

This product is intended for peptide enrichment and mass spectrometry analysis. To learn more about our Proteomics Kits and Services please answer a few questions for our Proteomics group.

Contact the CST Proteomics Group

    Product Information

    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. An alternate PTMScan® IAP Buffer Plus Detergent #9992, which may reduce nonspecific interactions, is available separately.

    Storage

    Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.

    Protocol

    Product Description

    PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology (CST) for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® Technology enables researchers to isolate, identify, and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity, providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur. For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/services/index.html.

    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).

    Lysine glutarylation events predominate in the mitochondria and mainly occur on metabolic proteins. Similar to malonylation and succinylation, lysine glutaryl transfer is likely mediated by a combination of non-enzymatic, thermodynamic, and stoichiometric mechanisms driving the reaction in the acidic environment of the mitochondria in the presence of promiscuous acetyltransferases (6). SirT5 and glutaryl-CoA dehydrogenase (GCDH) knockout mice exhibit elevated levels of glutarylated proteins as do mice on a diet high in tryptophan. SirT5 is likely the main de-glutarylase, as demonstrated by its activity on carbamoyl phosphate synthase 1 (CPS1), a heavily glutarylated protein of the urea cycle (7).
    For Research Use Only. Not For Use In Diagnostic Procedures.
    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    AcetylScan is a registered trademark of Cell Signaling Technology, Inc.
    MethylScan is a registered trademark of Cell Signaling Technology, Inc.
    PhosphoScan is a registered trademark of Cell Signaling Technology, Inc.
    UbiScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit our Trademark Information page.