Expression and purification of an engineered human endothelin receptor B in a monomeric form

Dokl Biochem Biophys. 2016 Mar;467(1):157-61. doi: 10.1134/S1607672916020216. Epub 2016 May 20.

Abstract

In humans, two endothelin receptors, ETa and ETb, are activated by three endogenous 21-mer cyclic peptides, ET-1, ET-2, and ET-3, which control various physiological processes, including vasoconstriction, vasodilation, and stimulation of cell proliferation. The first stage of this study it to produce a stable solubilized and purified receptor in a monodisperse state. This article is focused on the engineering, expression, purification, and characterization of the endothelin receptor B for subsequent structural and functional studies.

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Biphenyl Compounds / chemistry
  • Blotting, Western
  • Dipeptides / chemistry
  • Endothelin Receptor Antagonists / chemistry
  • Endothelins / chemistry
  • Genetic Engineering / methods
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Peptide Fragments / chemistry
  • Protein Denaturation
  • Protein Stability
  • Receptor, Endothelin B / chemistry*
  • Receptor, Endothelin B / genetics
  • Receptor, Endothelin B / isolation & purification*
  • Receptor, Endothelin B / metabolism
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Sf9 Cells
  • Temperature

Substances

  • Biphenyl Compounds
  • Dipeptides
  • EDNRB protein, human
  • Endothelin Receptor Antagonists
  • Endothelins
  • IFITM5 protein, human
  • IRL 2500
  • Membrane Proteins
  • Peptide Fragments
  • Receptor, Endothelin B
  • Recombinant Fusion Proteins
  • sovateltide