Cloning, purification and structure determination of the HIV integrase-binding domain of lens epithelium-derived growth factor

DOI: 10.1107/S2053230X18001553

Authors: Clare Hannon (Weatherall Institute of Molecular Medicine, University of Oxford; West Suffolk Hospital) , Abimael Cruz-Migoni (Weatherall Institute of Molecular Medicine, University of Oxford) , Olga Platonova (University of Oxford) , Robin L. Owen (Diamond Light Source) , Joanne E. Nettleship (The Oxford Protein Production Facility, Research Complex at Harwell, Rutherford Appleton Laboratory; University of Oxford) , Ami Miller (University of Oxford) , Stephen B. Carr (Weatherall Institute of Molecular Medicine, University of Oxford) , Gemma Harris (Research Complex at Harwell) , Terence H. Rabbitts (Weatherall Institute of Molecular Medicine, University of Oxford) , Simon E. V. Phillips (University of Oxford; Research Complex at Harwell)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acta Crystallographica Section F Structural Biology Communications , VOL 74 , PAGES 143 - 149

State: Published (Approved)
Published: March 2018
Diamond Proposal Number(s): 12346

Abstract: 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.

Journal Keywords: HIV integrase-binding domain; lens epithelium-derived growth factor; human immunodeficiency virus; domain swapping

Diamond Keywords: Human Immunodeficiency Virus (HIV); Viruses

Subject Areas: Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography

Added On: 01/03/2018 15:04

Documents:
pg5073.pdf

Discipline Tags:

Pathogens Infectious Diseases Health & Wellbeing Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)