Molecular cloning of murine CC CKR-4 and high affinity binding of chemokines to murine and human CC CKR-4.
We have cloned the murine homologue of human CC Chemokine Receptor-4 (CC CKR-4). In equilibrium competition binding assays performed in undifferentiated HL-60 cells transfected with human and murine CC CKR-4 cDNA, the IC50 values for the binding of [125I]macrophage inflammatory protein-1 alpha to human and murine CC CKR-4 were 14.5 +/- 9.0 nM and 10.1 +/- 3.0 nM, respectively, and the IC50 values for the binding of [125I]RANTES to human and murine CC CKR-4 were 9.3 +/- 3.0 nM and 5.7 +/- 2.6 nM, respectively. The cDNA clone for murine CC CKR-4 is 1531 bp, and the largest open reading frame encodes a protein of 360 amino acids that is 85% identical to human CC CKR-4. Murine CC CKR-4 was detected in the thymus and T-cell lines by Northern blot analysis. This first report of direct binding of chemokines to CC CKR-4 demonstrates that the highly homologous human and murine receptors have similar binding characteristics and tissue distribution.
Hoogewerf
A
A
Glaxo Institute for Molecular Biology, Geneva, Switzerland.
Black
D
D
Proudfoot
A E
AE
Wells
T N
TN
Power
C A
CA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D054407:Chemokine CCL4; D018946:Chemokine CCL5 / Q000378:metabolism; D003001:Cloning, Molecular; D015870:Gene Expression; D015723:Gene Library; D018922:HL-60 Cells; D006801:Humans; D007700:Kinetics; D008214:Lymphocytes / Q000378:metabolism*; D019402:Macrophage Inflammatory Proteins; D051379:Mice; D008969:Molecular Sequence Data; D015846:Monokines / Q000378:metabolism; D009928:Organ Specificity; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D018121:Receptors, Cytokine / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000378:metabolism; D013379:Substrate Specificity; D013950:Thymus Gland / Q000378:metabolism*; D014162:Transfection
C097843:CC cytokine receptor-4; D054407:Chemokine CCL4; D018946:Chemokine CCL5; D019402:Macrophage Inflammatory Proteins; D015846:Monokines; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D011994:Recombinant Proteins