Solubilization of the chemokine receptor CXCR4.
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.
Staudinger
R
R
Department of Pathology, Department of Neurology, Department of Medicine, New York University, School of Medicine, 423 East 23rd Street, New York, New York 10010, USA.
Bandres
J C
JC
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000378:metabolism; D002788:Cholesterol Esters / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D005960:Glucosides / Q000494:pharmacology; D015699:HIV Envelope Protein gp120 / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes / Q000378:metabolism; D007700:Kinetics; D011233:Precipitin Tests; D011485:Protein Binding; D019718:Receptors, CXCR4 / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*
C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D002788:Cholesterol Esters; D003902:Detergents; D005960:Glucosides; D015699:HIV Envelope Protein gp120; D007457:Iodine Radioisotopes; D019718:Receptors, CXCR4; D011994:Recombinant Proteins; C040358:dodecyl maltoside; C013440:cholesteryl succinate