Back to Search
Start Over
Role of the tryptophan residue in the vicinity of the catalytic center of exonuclease III family AP endonucleases: AP site recognition mechanism
- Source :
- Nucleic Acids Research
- Publication Year :
- 2006
- Publisher :
- Oxford University Press (OUP), 2006.
-
Abstract
- The mechanisms by which AP endonucleases recognize AP sites have not yet been determined. Based on our previous study with Escherichia coli exonuclease III (ExoIII), the ExoIII family AP endonucleases probably recognize the DNA-pocket formed at an AP site. The indole ring of a conserved tryptophan residue in the vicinity of the catalytic site presumably intercalates into this pocket. To test this hypothesis, we constructed a series of mutants of ExoIII and human APE1. Trp-212 of ExoIII and Trp-280 of APE1 were critical to the AP endonuclease activity and binding to DNA containing an AP site. To confirm the ability of the tryptophan residue to intercalate with the AP site, we examined the interaction between an oligopeptide containing a tryptophan residue and an oligonucleotide containing AP sites, using spectrofluorimetry and surface plasmon resonance (SPR) technology. The tryptophan residue of the oligopeptide specifically intercalated into an AP site of DNA. The tryptophan residue in the vicinity of the catalytic site of the ExoIII family AP endonucleases plays a key role in the recognition of AP sites.
- Subjects :
- Molecular Sequence Data
Article
Substrate Specificity
AP endonuclease
chemistry.chemical_compound
Catalytic Domain
DNA-(Apurinic or Apyrimidinic Site) Lyase
Escherichia coli
Genetics
AP site
Amino Acid Sequence
Binding site
Exonuclease III
Binding Sites
biology
Oligonucleotide
Tryptophan
DNA
DNA-(apurinic or apyrimidinic site) lyase
Exodeoxyribonucleases
Biochemistry
chemistry
Mutation
biology.protein
Peptides
Sequence Alignment
Subjects
Details
- ISSN :
- 13624962 and 03051048
- Volume :
- 34
- Database :
- OpenAIRE
- Journal :
- Nucleic Acids Research
- Accession number :
- edsair.doi.dedup.....19dae18d875c0205f00b23a730be88a1