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2-Hydrazinobenzothiazole-based etheno-adduct repair protocol (HERP): a method for quantitative determination of direct repair of etheno-bases.
Shivange, Gururaj; Kodipelli, Naveena; Anindya, Roy.
Afiliação
  • Shivange G; Department of Biotechnology, Indian Institute of Technology Hyderabad, Ordnance Factory Estate, Yeddumailaram, 502205 Hyderabad, India.
  • Kodipelli N; Department of Biotechnology, Indian Institute of Technology Hyderabad, Ordnance Factory Estate, Yeddumailaram, 502205 Hyderabad, India.
  • Anindya R; Department of Biotechnology, Indian Institute of Technology Hyderabad, Ordnance Factory Estate, Yeddumailaram, 502205 Hyderabad, India. Electronic address: anindya@iith.ac.in.
DNA Repair (Amst) ; 28: 8-13, 2015 Apr.
Article em En | MEDLINE | ID: mdl-25697729
Etheno-DNA adducts are mutagenic and lead to genomic instability. Enzymes belonging to Fe(II)/2-oxoglutarate-dependent dioxygenase family repair etheno-DNA adducts by directly removing alkyl chain as glyoxal. Presently there is no simple method to assess repair reaction of etheno-adducts. We have developed a rapid and sensitive assay for studying etheno-DNA adduct repair by Fe(II)/2-oxoglutarate-dependent dioxygenases. Using AlkB as model Fe(II)/2-oxoglutarate-dependent dioxygenases, we performed in vitro repair of etheno-adducts containing DNA and detected glyoxal by reacting with 2-hydrazinobenzothiazole which forms complex yellow color compound with distinct absorption spectrum with a peak absorption at 365 nm. We refer this method as 2-hydrazinobenzothiazole-based etheno-adduct repair protocol or HERP. Our novel approach for determining repair of etheno-adducts containing DNA overcomes several drawbacks of currently available radioisotope-based assay.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Espectrofotometria / Tiazóis / Adutos de DNA / Proteínas de Escherichia coli / Reparo do DNA / Oxigenases de Função Mista Idioma: En Ano de publicação: 2015 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Espectrofotometria / Tiazóis / Adutos de DNA / Proteínas de Escherichia coli / Reparo do DNA / Oxigenases de Função Mista Idioma: En Ano de publicação: 2015 Tipo de documento: Article