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result sql = Characterization of HIV Tat modifications using novel methyl-lysine-specific antibodies.
Characterization of HIV Tat modifications using novel methyl-lysine-specific antibodies.
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result sql = Modification-specific antibodies are important tools to examine the dynamics and functions of posttranslational protein modifications in cells. Here, we describe in detail the generation of polyclonal antibodies specific for mono-, di-, and trimethylated lysine 51 within the HIV transactivator Tat. Lysine 51 is a highly conserved residue located in the RNA-binding region of Tat and the target of lysine methyltransferases KMT1E (SETDB1) and KMT7 (Set7/9). Using affinity-purified methyl-specific antibodies of Tat, we find that cellular Tat is predominantly monomethylated at lysine 51, a modification enhanced by coexpression of KMT7.

Publication: Methods 2011 Jan ;53(1)::91-6.
Publication Date: 2011 Jan
Study Author(s): Pagans, Sara;Sakane, Naoki;Schnölzer, Martina;Ott, Melanie;
Institution: Gladstone Institute of Virology and Immunology, University of California, San Francisco, CA 94158, USA.
Shortcut link to this study:
Modification-specific antibodies are important tools to examine the dynamics and functions of posttranslational protein modifications in cells. Here, we describe in detail the generation of polyclonal antibodies specific for mono-, di-, and trimethylated lysine 51 within the HIV transactivator Tat. Lysine 51 is a highly conserved residue located in the RNA-binding region of Tat and the target of lysine methyltransferases KMT1E (SETDB1) and KMT7 (Set7/9). Using affinity-purified methyl-specific antibodies of Tat, we find that cellular Tat is predominantly monomethylated at lysine 51, a modification enhanced by coexpression of KMT7.
PMID: 20615470

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