Engineering a high-affinity methyl-CpG-binding protein

Helle F Jørgensen, Karen Adie, Pascal Chaubert, Adrian P Bird

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Core members of the MBD protein family (MeCP2, MBD1, MBD2 and MBD4) share a methyl-CpG-binding domain that has a specific affinity for methylated CpG sites in double-stranded DNA. By multimerizing the MDB domain of Mbd1, we engineered a poly-MBD protein that displays methyl-CpG-specific binding in vitro with a dissociation constant that is >50-fold higher than that of a monomeric MBD. Poly-MBD proteins also localize to methylated foci in cells and can deliver a functional domain to reporter constructs in vivo. We propose that poly-MBD proteins are sensitive reagents for the detection of DNA methylation levels in isolated native DNA and for cytological detection of chromosomal CpG methylation.
Original languageEnglish
Pages (from-to)e96
JournalNucleic Acids Research
Volume34
Issue number13
DOIs
Publication statusPublished - 2006

Keywords / Materials (for Non-textual outputs)

  • Animals
  • Cell Line
  • CpG Islands
  • DNA Methylation
  • DNA-Binding Proteins
  • Indicators and Reagents
  • Mice
  • Protein Engineering
  • Protein Structure, Tertiary
  • Recombinant Proteins

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