Expression and purification of NEIL3, a human DNA glycosylase homolog

Protein Expr Purif. 2009 Jun;65(2):160-4. doi: 10.1016/j.pep.2008.11.014. Epub 2008 Dec 13.

Abstract

The base excision repair (BER) pathway is mainly responsible for the repair of a vast number of non-bulky lesions produced by alkylation, oxidation or deamination of bases. DNA glycosylases are the key enzymes that recognize damaged bases and initiate BER by catalyzing the cleavage of the N-glycosylic bond between the base and the sugar. Many of the mammalian DNA glycosylases have been identified by a combination of biochemical and bioinformatics analysis. Thus, a mammalian family of three proteins (NEIL1, NEIL2 and NEIL3) that showed homology to the Escherichia coli Fpg/Nei DNA glycosylases was identified. Two of the proteins, NEIL1 and NEIL2 have been thoroughly characterized and shown to initiate BER of a diverse number of oxidized lesions. However, much less is known about NEIL3. The biochemical properties of NEIL3 have not been elucidated. This is mainly due to the difficulty in the expression and purification of NEIL3. Here, we describe the expression and partial purification of full-length human NEIL3 and the expression, purification and characterization of a truncated human core-NEIL3 (amino acids 1-301) that contains the complete E. coli Fpg/Nei-like domain but lacks the C-terminal region.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • Amino Acid Sequence
  • Cloning, Molecular
  • DNA Repair
  • Humans
  • Molecular Sequence Data
  • N-Glycosyl Hydrolases / biosynthesis*
  • N-Glycosyl Hydrolases / chemistry
  • N-Glycosyl Hydrolases / isolation & purification*

Substances

  • FLJ10858 protein, human
  • N-Glycosyl Hydrolases