Expanding on our offerings of long wavelength, highly sensitive
SIRT assays, AnaSpec, Eurogentec Group, is pleased to release 2 assay kits for
SIRT2, the SensoLyte®
520 FRET SIRT2 Assay Kit and the SensoLyte®
Green SIRT2 Assay Kit. The substrate found in the SensoLyte®
520 FRET SIRT2 Assay Kit employs a FRET substrate, and the peptide sequence
is derived from a human alpha-tubulin sequence surrounding the deacetylation
site of SIRT2. The SensoLyte®
Green SIRT2 Assay Kit provides a fluorogenic substrate with a human
p53 derived sequence. Compared to lower emitting substrates found in existing
commercial kits, these kits employ fluorimetric substrates that emit in the
green range of 520 nm. At this higher wavelength, the contribution of autofluorescence
These kits provide a convenient, two-step homogeneous procedure
for measuring sirtuin 2 activity and for screening enzyme inhibitors and activators.
In the first step, an acetylated substrate is incubated with sirtuin-containing
samples. In the second step, deacetylated substrate is cleaved by the sirtuin
developer, resulting in an increase of fluorescence that can be detected with
excitation at 490 nm and emission at 520 nm. Fluorescence produced is proportional
to SIRT2 activity.
Figure 1. Inhibition of SIRT2 activity by Ro-31-8220 as
measured with SensoLyte® 520 FRET SIRT2 Assay Kit (Cat# 72189).
Figure 2. Sensitivity of the SensoLyte®
520 FRET SIRT2 Assay Kit(Cat# 72189).
Histone deacetylases (HDACs) act
as transcriptional repressors of genes catalyzing the removal of acetyl groups
from a ε-N-acetyl lysine of histone.1 Sirtuins comprise a unique
class of nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases (class
III HDACs) that target multiple protein substrates to execute diverse biological
functions. Sirtuins catalyze a reaction that couples lysine deacetylation to
NAD hydrolysis, yielding O-acetyl-ADP-ribose and nicotinamide.2
Sirtuin 2 (SIRT2) belongs to the family of Sir2 (Silent Information
Regulator 2) proteins. Substrates for SIRT2 are not limited to histones, but
also include various transcription factors such as forkhead box (FOXO) transcription
factors (FOXO1, FOXO3a) that modulate metabolic, cell cycle and cell death related
pathways.3, 4 Other prominent substrates include the tumor suppressor
p53 and alpha-tubulin.5, 6 Human SIRT2 is a cytoplasmic protein that
increases in abundance during mitosis and regulates major events of cytokinesis.7
Recombinant human SIRT2, a
positive control included in the kits, is also
available as a separate item.
- Sterner, DE. et al. Microbiol. Mol. Biol. Rev. 64,
- Longo, V and Kennedy, B. Cell. 126, 257 (2006).
- Wang, F. et al. Aging Cell 6, 505 (2007).
- Jing, E. et al. Cell Metab. 6, 105 (2007).
- Jin, Y.-H. et al. Biochem. Biophys. Res. Commun. 368,
- Inoue, T. et al. Cell Cycle 6(9) (2007) .
- Dryden, S. et al. Mol. Cell. Biol. 23, 3173