In vitro evolution of an HIV integrase binding protein from a library of C-terminal domain γS-crystallin variants

Bioorg Med Chem Lett. 2012 Sep 1;22(17):5584-9. doi: 10.1016/j.bmcl.2012.07.008. Epub 2012 Jul 10.

Abstract

A protein without natural binding functions was engineered to bind HIV-1 integrase. Phage display selections applied a library of variants based on the C-terminal domain of the eye lens protein human γS-crystallin. Multiple loop regions were altered to encode libraries with ≈3.6 × 10(11) different variants. A crystallin variant, termed integrase binding protein-10 (IBP-10), inhibits integrase catalysis with nanomolar K(i) values. IBP-10 interacts with the integrase C-terminal domain and inhibits integrase substrate affinity. This allosteric mechanism allows IBP-10 to inhibit drug-resistant integrase variants. The results demonstrate the applicability of the crystallin scaffold for the discovery of binding partners and enzyme inhibitors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • HIV Infections / drug therapy
  • HIV Infections / virology
  • HIV Integrase / chemistry
  • HIV Integrase / metabolism*
  • HIV Integrase Inhibitors / chemistry
  • HIV Integrase Inhibitors / metabolism
  • HIV Integrase Inhibitors / pharmacology*
  • HIV-1 / drug effects
  • HIV-1 / enzymology*
  • Humans
  • Models, Molecular
  • Peptide Library
  • Protein Binding
  • Protein Engineering / methods*
  • Protein Structure, Tertiary
  • gamma-Crystallins / chemistry
  • gamma-Crystallins / genetics
  • gamma-Crystallins / pharmacology*

Substances

  • HIV Integrase Inhibitors
  • Peptide Library
  • gamma-Crystallins
  • HIV Integrase
  • p31 integrase protein, Human immunodeficiency virus 1