Acetyl-p53 (Lys382) Antibody

2525:

Western Blotting

Acetyl-p53 (Lys382) Antibody detects endogenous levels of p53 only when acetylated at lysine 382. This antibody does not cross-react with other acetylated proteins.

Polyclonal antibodies are produced by immunizing animals with a synthetic acetylated peptide corresponding to residues surrounding Lys382 of human p53. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with both trichostatin A #9950 (400 nM for 24 hours), and doxorubicin (0.5 µM for 24 hours) using Acetyl-p53 (Lys382) Antibody alone (A), antibody preincubated with a non-acetylated Lys382 peptide (B), or antibody preincubated with an acetylated Lys382 peptide (C).

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated, trichostatin A-treated #9950 (400 nM for 24 hours), doxorubicin-treated (0.5 µM for 24 hours), or both, using Acetyl-p53 (Lys382) Antibody (top) or p53 Antibody #2524 (bottom).

The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (1). p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro (2,3). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to a reduced interaction between p53 and its negative regulator, the oncoprotein MDM2 (4). MDM2 inhibits p53 accumulation by targeting it for ubiquitination and proteasomal degradation (5,6). p53 can be phosphorylated by ATM, ATR, and DNA-PK at Ser15 and Ser37. Phosphorylation impairs the ability of MDM2 to bind p53, promoting both the accumulation and activation of p53 in response to DNA damage (4,7). Chk2 and Chk1 can phosphorylate p53 at Ser20, enhancing its tetramerization, stability, and activity (8,9). p53 is phosphorylated at Ser392 in vivo (10,11) and by CAK in vitro (11). Phosphorylation of p53 at Ser392 is increased in human tumors (12) and has been reported to influence the growth suppressor function, DNA binding, and transcriptional activation of p53 (10,13,14). p53 is phosphorylated at Ser6 and Ser9 by CK1δ and CK1ε both in vitro and in vivo (13,15). Phosphorylation of p53 at Ser46 regulates the ability of p53 to induce apoptosis (16). Acetylation of p53 is mediated by p300 and CBP acetyltransferases. Inhibition of deacetylation suppressing MDM2 from recruiting HDAC1 complex by p19 (ARF) stabilizes p53. Acetylation appears to play a positive role in the accumulation of p53 protein in stress response (17). Following DNA damage, human p53 becomes acetylated at Lys382 (Lys379 in mouse) in vivo to enhance p53-DNA binding (18). Deacetylation of p53 occurs through interaction with the SIRT1 protein, a deacetylase that may be involved in cellular aging and the DNA damage response (19).

The histone acetyltransferases p300 and PCAF can acetylate p53 in vitro at Lys382 and Lys320, respectively (17). Lys382 becomes acetylated in vivo following DNA damage to allow enhanced p53-DNA binding (18).

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4. Shieh, S.Y. et al. (1997) Cell 91, 325-334.
5. Chehab, N.H. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 13777-13782.
6. Honda, R. et al. (1997) FEBS Lett. 420, 25-27.
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8. Shieh, S.Y. et al. (1999) EMBO J. 18, 1815-1823.
9. Hirao, A. et al. (2000) Science 287, 1824-1827.
10. Hao, M. et al. (1996) J. Biol. Chem. 271, 29380-29385.
11. Lu, H. et al. (1997) Mol. Cell. Biol. 17, 5923-5934.
12. Ullrich, S.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 5954-5958.
13. Kohn, K.W. (1999) Mol. Biol. Cell 10, 2703-2734.
14. Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-2539.
15. Knippschild, U. et al. (1997) Oncogene 15, 1727-1736.
16. Oda, K. et al. (2000) Cell 102, 849-862.
17. Ito, A. et al. (2001) EMBO J. 20, 1331-1340.
18. Sakaguchi, K. et al. (1998) Genes Dev. 12, 2831-2841.
19. Solomon, J.M. et al. (2006) Mol. Cell. Biol. 26, 28-38.
20. Gu, W. and Roeder, R.G. (1997) Cell 90, 595-606.
21. Sakaguchi, K. et al. (1998) Genes Dev. 12, 2831-2841.

• Kumazawa, T. et al. (2011) J Biol Chem 286, 20861-9. Applications: Western Blotting

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