1. Cloning, purification and structure determination of the HIV integrase-binding domain of lens epithelium-derived growth factor
- Author
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Hannon, Clare, Cruz-Migoni, Abimael, Platonova, Olga, Owen, Robin L., Nettleship, Joanne E., Miller, Ami, Carr, Stephen B., Harris, Gemma, Rabbitts, Terence H., and Phillips, Simon E. V.
- Subjects
Models, Molecular ,human immunodeficiency virus ,Protein Conformation ,HIV Integrase ,HIV integrase-binding domain ,Crystallography, X-Ray ,digestive system ,digestive system diseases ,Research Communications ,Catalytic Domain ,Humans ,Amino Acid Sequence ,domain swapping ,Crystallization ,lens epithelium-derived growth factor ,Adaptor Proteins, Signal Transducing ,Transcription Factors - Abstract
The HIV integrase-binding domain (IBD) of lens epithelium-derived growth factor has been cloned, purified and crystallized, and its structure has been solved. IBD forms an unusual domain-swapped dimer that assembles into octamers in the crystal., Lens epithelium-derived growth factor (LEDGF)/p75 is the dominant binding partner of HIV-1 integrase in human cells. The crystal structure of the HIV integrase-binding domain (IBD) of LEDGF has been determined in the absence of ligand. IBD was overexpressed in Escherichia coli, purified and crystallized by sitting-drop vapour diffusion. X-ray diffraction data were collected at Diamond Light Source to a resolution of 2.05 Å. The crystals belonged to space group P21, with eight polypeptide chains in the asymmetric unit arranged as an unusual octamer composed of four domain-swapped IBD dimers. IBD exists as a mixture of monomers and dimers in concentrated solutions, but the dimers are unlikely to be biologically relevant.
- Published
- 2018