Solubilization of the chemokine receptor CXCR4

Biochem Biophys Res Commun. 2000 Jul 21;274(1):153-6. doi: 10.1006/bbrc.2000.3109.

Abstract

The chemokine receptor CXCR4 was solubilized from the human T-cell line CEM by using the detergent n-dodecyl-beta-maltoside (DDM) and cholesteryl hemisuccinate ester (CHS). Binding studies with (125)I-SDF-1alpha revealed a dissociation constant of 5.33 nM and a receptor density (B(max)) of 2.68 pmol/mg in CEM membranes at 4 degrees C. The affinity of solubilized CXCR4 for SDF-1alpha was identical to membrane-bound CXCR4. Binding of gp120 to solubilized CXCR4 was demonstrated by coprecipitation of gp120 with anti-CXCR4 antibodies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line
  • Cell Membrane / metabolism
  • Chemokine CXCL12
  • Chemokines, CXC / metabolism
  • Cholesterol Esters / pharmacology
  • Detergents / pharmacology
  • Glucosides / pharmacology
  • HIV Envelope Protein gp120 / metabolism
  • HIV-1 / metabolism
  • Humans
  • Iodine Radioisotopes / metabolism
  • Kinetics
  • Precipitin Tests
  • Protein Binding
  • Receptors, CXCR4 / isolation & purification
  • Receptors, CXCR4 / metabolism*
  • Recombinant Proteins / metabolism
  • T-Lymphocytes / metabolism*

Substances

  • CXCL12 protein, human
  • Chemokine CXCL12
  • Chemokines, CXC
  • Cholesterol Esters
  • Detergents
  • Glucosides
  • HIV Envelope Protein gp120
  • Iodine Radioisotopes
  • Receptors, CXCR4
  • Recombinant Proteins
  • dodecyl maltoside
  • cholesteryl succinate