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

Acta Crystallogr F Struct Biol Commun. 2018 Mar 1;74(Pt 3):143-149. doi: 10.1107/S2053230X18001553. Epub 2018 Feb 26.

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.

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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / chemistry*
  • Adaptor Proteins, Signal Transducing / isolation & purification
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Amino Acid Sequence
  • Catalytic Domain
  • Crystallization
  • Crystallography, X-Ray
  • HIV Integrase / chemistry*
  • HIV Integrase / metabolism*
  • Humans
  • Models, Molecular
  • Protein Conformation
  • Transcription Factors / chemistry*
  • Transcription Factors / isolation & purification
  • Transcription Factors / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • PSIP1 protein, human
  • Transcription Factors
  • HIV Integrase