Crystal structure of the DNA repair enzyme endonuclease-VIII (Nei) from E. coli (E2Q) in complex with AP-site containing DNA substrate

OPEN FORM OF 1'-2'-DIDEOXYRIBOFURANOSE-5'-PHOSPHATE

Active site plasticity of endonuclease VIII: Conformational changes compensating for different substrates and mutations of critical residues

DNA glycosylase/AP lyase Nei, DNA-apurinic or apyrimidinic site, lyase Nei

1.8M ammonium sulfate, 0.1M sodium acetate pH 4.6-5.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K

SEQUENCING AND CRYSTALLOGRAPHY CONFIRM THE SEQUENCE AS THR A 34, THR B 34, ARG A 112, ARG B 112.

Data processing indicated that the crystal was twinned but the exact nature of the twining was not clear. Treatment of the data with the twinning utility of CNS indicated unequal (major and minor) twinning fractions, but a refinement of the initial model, ignoring the twinning and considering only the major twinning fraction, gave better structural results, with relatively reasonable final R and Rfree.

Endonuclease VIII, oxidative damage, DNA repair, base excision, covalent intermediate, reaction mechanism, HYDROLASE-DNA COMPLEX