Expression and Purification of the Human Xeroderma Pigmentosum F (XPF/ERCC1) Complex in Bacterial Cells for Protein Binding Assays and Crystallographic Studies

Abstract

Mutations in dna repair enzymes of the XP complementation group, XPA‐XPG, leads to a number of genetic disorders such as Xeroderma Pigmentosum, Cockayne's Syndrome, and Trichothiodystrophy. The structure specific dna repair endonuclease responsible for the 5′ incision during dna repair, XPF, is involved in homologous recombination that assists in removing interstrand cross‐link. XP‐F is an autosomal recessive disease characterized by hypersensitivity of the skin to sunlight followed by high incidence of skin cancer and frequent neurologic abnormalities. I expressed and purified human recombinant XPF and ERCC1 proteins in E.Coli Rosetta and BL21 cells, these results will be scaled up for use in interaction and crystallographic studies with proteins of the XP family.