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Functional and biochemical analysis of the cloned Duffy antigen: identity
with the red blood cell chemokine receptor
K Neote, JY Mak, LF Kolakowski and TJ Schall
Department of Immunology, Genentech Inc, South San Francisco, CA.
The Duffy blood group antigen has been postulated to be a receptor on red
blood cells (RBCs) for the malarial parasite Plasmodium vivax and a
promiscuous receptor for the chemokine superfamily of inflammatory
proteins. Recently, the Duffy antigen glycoprotein D cDNA has been cloned
(Chaudhuri et al: Proc Natl Acad Sci USA 90:10793, 1993). We have analyzed
the binding properties of the cloned Duffy antigen. Duffy- antigen cDNAs
expressed in human embryonic kidney cells produced cell- surface proteins
that reacted with two known anti-Duffy monoclonal antibodies. Direct ligand
binding and displacement experiments using recombinant chemokine proteins
also show that the cloned Duffy protein is the RBC chemokine receptor.
Radiolabeled chemokines of both the C-C (RANTES and MCP-1) and C-X-C (IL-8
and MGSA/gro) subclasses bound reversibly to transfected cells with
dissociation constants in the nanomolar range. Chemokines of either class
displaced heterologous chemokines, indicating that they were competing for
a single site on the transfected cells. Although the chemokines bound to
the transfected cells with high affinity, there was no evidence for signal
transduction, as measured by transient increases in intracellular calcium
ion concentration, through the Duffy antigen/RBC chemokine receptor in
transfected cells. Lastly, we have performed a computer analysis on the
amino acid structure of the Duffy antigen/RBC chemokine receptor. Although
the cloned Duffy antigen has been postulated to be a
nine-transmembrane-spanning receptor, our analysis suggests that the
molecule most likely belongs to the seven-transmembrane-spanning receptor
superfamily and is therefore similar to other chemokine receptors
previously identified.
Volume 84,
Issue 1,
pp. 44-52,
07/01/1994
Copyright © 1994 by The American Society of Hematology
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