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.
Copyright 2000 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Cell Line
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Cell Membrane / metabolism
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Chemokine CXCL12
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Chemokines, CXC / metabolism
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Cholesterol Esters / pharmacology
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Detergents / pharmacology
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Glucosides / pharmacology
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HIV Envelope Protein gp120 / metabolism
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HIV-1 / metabolism
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Humans
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Iodine Radioisotopes / metabolism
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Kinetics
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Precipitin Tests
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Protein Binding
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Receptors, CXCR4 / isolation & purification
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Receptors, CXCR4 / metabolism*
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Recombinant Proteins / metabolism
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T-Lymphocytes / metabolism*
Substances
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CXCL12 protein, human
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Chemokine CXCL12
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Chemokines, CXC
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Cholesterol Esters
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Detergents
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Glucosides
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HIV Envelope Protein gp120
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Iodine Radioisotopes
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Receptors, CXCR4
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Recombinant Proteins
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dodecyl maltoside
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cholesteryl succinate