Quantitative LC-MS/MS analysis of 5-hydroxymethyl-2'-deoxyuridine to monitor the biological activity of J-binding protein.
Anal Biochem
; 610: 113930, 2020 12 01.
Article
em En
| MEDLINE
| ID: mdl-32866463
ABSTRACT
Base J replaces 1% of thymine in most kinetoplastid flagellates, and is implicated in transcription regulation. Base J is synthesized in two steps:
first, a thymine base in DNA is converted to 5-hydroxymethyluracil by J-binding proteins (JBP1, JBP2); secondly, a glucosyl transferase glycosylates the 5-hydroxymethyluracil to form base J. Here, we present a highly sensitive and selective LC-MS/MS method to quantify the in vitro JBP1 activity on synthetic oligonucleotide substrates. The method demonstrated successful to support biochemical studies of JBPs and can be used as a template for additional JBP activity studies or for inhibitor screening in the future.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Timidina
/
Proteínas de Protozoários
/
Cromatografia Líquida de Alta Pressão
/
Proteínas de Ligação a DNA
/
Espectrometria de Massas em Tandem
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Holanda