期刊
BIOMOLECULAR NMR ASSIGNMENTS
卷 4, 期 1, 页码 5-8出版社
SPRINGER
DOI: 10.1007/s12104-009-9196-y
关键词
APE1; Ref-1; APEX1; Apurinic/apyrimidinic endonuclease; DNA base excision repair; NMR chemical shift assignments
资金
- National Institutes of Health [R01-GM72711]
- University of Maryland Marlene
- Stewart Greenebaum Cancer Center
- NIH [T32-GM066706, S10-RR10441, S10-RR15741, S10-RR16812, S10-RR23447]
- NSF [DBI 0115795]
Apurinic/apyrimidinic endonuclease 1 (APE1 or Ref-1) is the major enzyme in mammals for processing abasic sites in DNA. These cytotoxic and mutagenic lesions arise via spontaneous rupture of the base-sugar bond or the removal of damaged bases by a DNA glycosylase. APE1 cleaves the DNA backbone 5' to an abasic site, giving a 3'-OH primer for repair synthesis, and mediates other key repair activities. The DNA repair functions are essential for embryogenesis and cell viability. APE1-deficient cells are hypersensitive to DNA-damaging agents, and APE1 is considered an attractive target for inhibitors that could potentially enhance the efficacy of some anti-cancer agents. To enable an important new method for studying the structure, dynamics, catalytic mechanism, and inhibition of APE1, we assigned the chemical shifts (backbone and C-13(beta)) of APE1 residues 39-318. We also report a protocol for refolding APE1, which was essential for achieving complete exchange of backbone amide sites for the perdeuterated protein.
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