| Products/Services Used | Details | Operation |
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| Protein Electrophoresis and Western | Fractions containing 6xH-SIRT7 were identified by SDS-PAGE (12% acrylamide gel) and combined, supplemented with DTT (2 mM) and TEV protease (cat. # Z03030, GenScript), and dialyzed against buffer C (50 mM K2HPO4, 50 mM NaCl, 1 mM DTT, pH 7.5) at 4 °C overnight.Then, ~0.25 µg protein were analyzed by SDS-PAGE (SurePAGE 4-12% gels, Genscript cat. # M00654, ran at 100 V and stained with Coomasie blue) together with a set of BSA standard dilutions (TFS, cat. # 23209), | Get A Quote |
Chromatin-modifying enzymes target distinct residues within histones to finetune gene expression profiles. SIRT7 is an NAD+-dependent deacylase often deregulated in cancer, which deacetylates either H3 lysine 36 (H3K36) or H3K18 with high specificity within nucleosomes. Here, we report structures of nucleosome-bound SIRT7, and uncover the structural basis of its specificity towards H3K36 and K18 deacylation, combining a mechanism-based cross-linking strategy, cryo-EM, and enzymatic and cellular assays. We show that the SIRT7 N-terminus represents a unique, extended nucleosome-binding domain, reaching across the nucleosomal surface to the acidic patch. The catalytic domain binds at the H3-tail exit site, engagin... More
