Transplantation of a 17-amino acid alpha-helical DNA-binding domain into an antibody molecule confers sequence-dependent DNA recognition

K E McLane, D R Burton, P Ghazal

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Recombinant antibodies capable of sequence-specific interactions with nucleic acids represent a class of DNA- and RNA-binding proteins with potential for broad application in basic research and medicine. We describe the rational design of a DNA-binding antibody, Fab-Ebox, by replacing a variable segment of the immunoglobulin heavy chain with a 17-amino acid domain derived from TFEB, a class B basic helix-loop-helix protein. DNA-binding activity was studied by electrophoretic mobility-shift assays in which Fab-Ebox was shown to form a specific complex with DNA containing the TFEB recognition motif (CACGTG). Similarities were found in the abilities of TFEB and Fab-Ebox to discriminate between oligodeoxyribonucleotides containing altered recognition sequences. Comparable interference of binding by methylation of cytosine residues indicated that Fab-Ebox and TFEB both contact DNA through interactions along the major groove of double-stranded DNA. The results of this study indicate that DNA-binding antibodies of high specificity can be developed by using the modular nature of both immunoglobulins and transcription factors.
Original languageEnglish
Pages (from-to)5214-8
Number of pages5
JournalProceedings of the National Academy of Sciences (PNAS)
Issue number11
Publication statusPublished - 1995

Keywords / Materials (for Non-textual outputs)

  • Amino Acid Sequence
  • Antibodies
  • Base Sequence
  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
  • Consensus Sequence
  • DNA
  • DNA Primers
  • DNA-Binding Proteins
  • Helix-Loop-Helix Motifs
  • Immunoglobulin Fab Fragments
  • Kinetics
  • Molecular Sequence Data
  • Neoplasm Proteins
  • Oligonucleotide Probes
  • Polymerase Chain Reaction
  • Protein Structure, Secondary
  • RNA-Binding Proteins
  • Recombinant Proteins
  • Substrate Specificity
  • Transcription Factors


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