| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | Antibodies against citrulline-containing peptides were raised in rabbits by the BioGenes company using the peptide sequences SFASDPILY-CIT-PVAVALDTKGGC and ASFI-Cit-KASDVHEVRKVLGEGGC for R106Cit and R246Cit respectively. Peptides were generated, quantified and confirmed by mass spectrometry by Genscript. Rabbits were immunized with carrier-conjugated peptide followed by three 3 booster injections after 14, 28 and 42 days. Affinity purification of antisera were performed by coupling the citrullinated peptides or their wild-type counterparts to SulfoLink Coupling Gel (PIERCE, 20401) agarose beads. The antisera were passed first through the column with citrullinated peptides and then through column with the wild-type peptide to remove residual antibodies recognizing the wild-type peptide. | Get A Quote |
CHD3 and CHD4 are mutually exclusive ATPase subunits of the Nucleosome Remodelling and Deacetylation (NuRD) complex that regulates gene expression. CHD4 is essential for growth of multiple patient derived melanoma xenografts and for breast cancer. Here we show that CHD4 regulates expression of PADI1 (Protein Arginine Deiminase 1) and PADI3 in multiple cancer cell types modulating citrullination of three arginines of the allosterically-regulated glycolytic enzyme pyruvate kinase M2 (PKM2). Citrullination reprograms cross-talk between PKM2 ligands lowering its sensitivity to the inhibitors Tryptophan, Alanine and Phenylalanine and promoting activation by Serine. Citrullination thus bypasses normal physiological r... More
