Protocol for an in vitro assay to study HIV-1 Tat methylation

被引：0
|
作者：
Boehm, Daniela [1 ,2 ]
Kaehlcke, Katrin [1 ,4 ]
Schnoelzer, Martina [5 ]
Ott, Melanie [1 ,2 ,3 ]
机构：
[1] Univ Calif San Francisco, Gladstone Inst Virol, San Francisco, CA 94158 USA
[2] Univ Calif San Francisco, Dept Med, San Francisco, CA 94143 USA
[3] Chan Zuckerberg Biohub, San Francisco, CA 94158 USA
[4] Univ Med Ctr Freiburg, D-79106 Freiburg, Germany
[5] German Canc Res Ctr, Funct Proteome Anal, D-69120 Heidelberg, Germany
来源：
STAR PROTOCOLS | 2023年 / 4卷 / 04期
基金：
美国国家卫生研究院;
关键词：
Cell Biology; Molecular Biology; Protein Biochemistry;
D O I：
10.1016/j.xpro.2023.102687
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A critical virus-encoded regulator of HIV-1 transcription is the Tat protein, which is required to potently activate transcription. Tat is regulated by a wide variety of post-translational modifications. This protocol describes an in vitro assay to study Tat methylation. We describe steps for incorporation of radioactive methyl groups into Tat protein, visualization by gel analysis, Coomassie blue stain, gel drying, and detection by autoradiography. This protocol can also be used to assess methylation in other proteins such as histones.For complete details on the use and execution of this protocol, please refer to Boehm et al. (2023).1
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页数：9
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