FLUOR DE LYS Substrates & Accessories
FLUOR DE LYS substrates form the core of the FLUOR DE LYS Substrate/Developer system for measuring HDAC activity. These include the original FLUOR DE LYS Substrate (BML-KI104), which incorporates a single acetylated lysine and is the most versatile of the series. It is acted upon by class I, class II and sirtuin deacetylases, and is suitable for measuring activity from extracts, immunoprecipitated or recombinant protein, and intact cells. Further substrates have been developed based on short peptide sequences (4-6 amino acids) from lysine acetylation sites in histones H3 and H4, and p53. While no substrate is absolutely specific for a given enzyme, the following table provides our recommendations for the optimal substrates to use with a given enzyme:
Substrate |
HDAC |
|
|
|
|
|
|
|
Sirtuin |
|
|
|
|
1 |
2 |
3 |
6 |
8 |
10 |
11 |
HeLa nuclear extract |
1 |
2 |
3 |
5 |
FLUOR DE LYS |
+++ |
++ |
++++ |
++ |
+ |
++++ |
++ |
++ |
+ |
+ |
+ |
+ |
FLUOR DE LYS H4-AcK16 |
+++ |
+++ |
++++ |
+++++ |
+++ |
n.d. |
+++ |
+++ |
++++ |
+++ |
+ |
+++ |
FLUOR DE LYS HDAC8 |
++++ |
+++++ |
++++ |
+++++ |
+++++ |
n.d. |
+++++ |
+++++ |
+++ |
++++ |
+++ |
+++++ |
FLUOR DE LYS SIRT1 |
+++++ |
+++++ |
+++++ |
+++++ |
+ |
+++++ |
+++++ |
+++++ |
+++++ |
+++++ |
+++ |
+++++ |
FLUOR DE LYS SIRT2 |
+++ |
++++ |
++++ |
++++ |
++ |
+ |
+ |
++++ |
++ |
+++++ |
+++++ |
++++ |
FLUOR DE LYS Substrate concentration |
5µM |
5µM |
50µM |
50µM |
25µM |
25µM |
5µM |
25µM |
25/500µM |
25µM |
10µM |
500µM |
Table legend:
Substrate preferences for a number of HDACs and sirtuins. The table indicates the substrate preferences for the indicated deacetylase among a panel of FLUOR DE LYS substrates based on short stretches of human histones H3, H4 and p53 sequence. The concentration of each substrate is indicated in the last row. For sirtuins the concentration of the co-substrate NAD+ was 500μM. Class IIa enzymes (HDACs 4, 5, 7 and 9) were not included because they exhibit very little activity with standard peptide substrates (see Lahm et al.; PNAS 104, 17335 (2007)).
n.d. indicates not determined.