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Full-Text Articles in Medical Molecular Biology
Interaction Of Fluorescent Probes With Sirtuin Proteins, James Fusco, Brian P Weiser
Interaction Of Fluorescent Probes With Sirtuin Proteins, James Fusco, Brian P Weiser
Rowan-Virtua Research Day
Sirtuins are a class of proteins belonging to the Sir2 (Silencing information regulator 2) family of NAD+ dependent protein lysine deacylases. Different Isoforms (SIRT1-SIRT7) differ in their specific deacylase activity and cellular location. They have roles in DNA repair, glucose metabolism, and cellular proliferation which make them highly desirable targets for carcinoma therapeutics. We previously used 1-aminoanthracene’s (AMA) fluorescent properties when bound with SIRT2 (Kd of 37 μM) to develop a high-throughput screen to identify novel ligands that inhibit SIRT2’s enzymatic activities. We hope to reveal other potential probes for future high-throughput screening with all the sirtuin isotopes. 1-AMA’s fluorescence …
Substrate-Dependent Modulation Of Sirt2 By A Fluorescent Probe, 1-Aminoanthracene, David Bi, Prashit Parikh, Jie Yang, Brian P Weiser
Substrate-Dependent Modulation Of Sirt2 By A Fluorescent Probe, 1-Aminoanthracene, David Bi, Prashit Parikh, Jie Yang, Brian P Weiser
Rowan-Virtua Research Day
Sirtuin isoform 2 (SIRT2) is an enzyme that catalyzes the removal of acyl groups from lysine residues. SIRT2’s catalytic domain has a hydrophobic tunnel where its substrate acyl groups bind. Here, we report that the fluorescent probe 1-aminoanthracene (AMA) binds within SIRT2’s hydrophobic tunnel in a substrate-dependent manner. AMA’s interaction with SIRT2 was characterized by its enhanced fluorescence upon protein binding (>10-fold). AMA interacted weakly with SIRT2 alone in solution (Kd = 37 μM). However, when SIRT2 was equilibrated with a decanoylated peptide substrate, AMA’s affinity for SIRT2 was enhanced ∼10-fold (Kd = 4μM). The peptide’s decanoyl chain and …