SPR imaging for label - free multiplexed analyses of DNA N - glycosylase interactions with damaged DNA duplexes

  • Corne C
  • Fiche J
  • Gasparutto D
 et al. 
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Abstract

See, stats, and : https : / / www . researchgate . net / publication / 51419530 SPR - free analyses - glycosylase with Article DOI : 10 . 1039 / b801086b : PubMed CITATIONS 24 READS 49 10 , including : Some : electronic Digital Didier Atomic 161 , 144 SEE Arnaud Atomic 78 , 147 SEE Roberto Atomic 138 , 873 SEE Thierry 126 , 540 SEE All . The . PAPER www . rsc . org / analyst | The Analyst Base excision repair (BER) is the major mechanism for the correction of damaged nucleobases resulting from the alkylation and oxidation of DNA . The first step in the BER pathway consists of excision of the abnormal base by several specific DNA N - glycosylases . A decrease in BER activity was found to be related to an increased risk of carcinogenesis and aging . To investigate BER activities we set up a new device for DNA repair analysis based on surface plasmon resonance imaging (SPRi) . Oligonucleotides bearing an abnormal nucleoside , namely 8 - oxo - 7 , 8 - dihydro - 2 - deoxyguanosine and (5 S) - 5 , 8 - cyclopurine - 2 - deoxynucleoside , were grafted by a pyrrole electro - copolymerization process on a glass prism coated with a gold layer . The latter label - free DNA sensor chip permits the detection of N - glycosylase / AP - lyase activity as well as the binding of repair proteins to DNA damage without cleavage activity . Thus , the Fapy DNA N - glycosylase (Fpg) protein is shown as expected to bind and then cleave its natural substrate , namely 8 - oxo - 7 , 8 - dihydro - guanine , together with the resulting abasic site . Using the current SPR imaging - based DNA array we observed an original binding activity of Fpg towards the (5 S) - 5 , 8 - cyclodAdenosine residue . These results altogether show that SPR imaging may be used to simultaneously and specifically detect recognition and excision of several damaged DNA nucleobases , and constitutes an interesting technique to screen inhibitors of DNA repair proteins .

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Authors

  • Christelle Corne

  • Jean-Bernard Fiche

  • Didier Gasparutto

  • Valérie Cunin

  • Emmanuel Suraniti

  • Arnaud Buhot

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