Based on the excellent High-Affi™ technology, Creative Biolabs has the capability of providing highly specific pan anti-glutaryllysine antibody and site-specific anti-glutarylated protein antibodies production services. These antibodies are produced by immunizing animals with glutarylated BSA or KLH or with a synthetic glutaryl peptide and purified by affinity chromatography. Besides, we can offer the scFv, Fab, full-length antibodies and single domain antibodies based on our phage display technology. The pan anti-glutaryllysine antibody specifically recognizes proteins with glutarylated lysine residues without species limitation, and site-specific anti-glutarylated antibodies recognize site-specific glutarylated peptide. They do not cross-react with non-modified lysine residues, unmodified peptides, malonylated peptides and succinyl peptides. The antibodies have been widely used in proteomic screening to lysine glutarylated substrates.
Glutarylation is a newly identified and validated post-translational modification (PTM) by mass spectrometry (MS) and biochemical methods in 2014. It is the addition of five-carbon dicarboxylates to lysine residues with structurally similar to malonylation and succinylation. Lysine glutarylation is highly conserved and widely distributed from prokaryotes to eukaryotes. It is also a reversible and dynamic process that is transferred by the enzyme p300 in vitro and removed by NAD+ dependent deacetylase, sirtuin 5 (SIRT5) both in vitro and in vivo. Moreover, it is probable that glutaryl-lysine is the largest substrate that Sirt5 can accommodate, as adipoyl-lysine is not cleaved by Sirt5 under in vitro conditions.
Fig. 1. Structures of glutaryl-lysine, succinyl-lysine and malonyl-lysine. (Tan M, et al. 2014)
Glutarylation is abundant in metabolic enzymes and mitochondrial proteins. Like other acylation, glutarylation was regulated by the acyl-donor glutaryl-CoA which is produced in mitochondria as a result of the catabolism of tryptophan and lysine.
It is proposed that glutarylation could affect transcriptional gene expression through mechanism similar to malonylation and succinylation. Carbamoyl phosphate synthase 1 (CPS1), the rate-limiting enzyme in the urea cycle, was the first identified glutarylated protein. They demonstrated that the activities of CPS1 was regulated by glutarylation with improved glutarylation suppresses CPS1 enzymatic activity. As a consequence, nitrogen balance maintained by CPS1 was affected and then lead to the metabolic disorder glutaric acidemia (GA). Knockout or inactivated SIRT5 leads to the inhibition of G6PD with high-level glutarylation, thereby decreasing the production of NADPH, lowering GSH, impairing the ability to scavenge ROS, and increasing cellular suffering from oxidative stress.
Fig. 2. Glutarylation of CPS1 is regulated by deacetylase SIRT5. (Tan M, et al. 2014)
With advanced MS technological and high-quality antibodies, the more physiological and pathological roles of glutarylation will be uncovered soon. Creative Biolabs has many years of experience in the field of PTM antibody discovery and production. The professional experts will provide our customers with the best and comprehensive guaranteed products and services.
Creative Biolabs can provide a comprehensive list of PTM-specific antibody production services of your choice.