Interleukin-8 Receptor 
THE ROLE OF THE CARBOXYL TERMINUS IN SIGNAL TRANSDUCTION (*)
- Adit Ben-Baruch(1)(§),
- Kathleen M. Bengali(2),
- Arya Biragyn(3),
- Jim J. Johnston(4),
- Ji-Ming Wang(2),
- Jin Kim(5),
- Anan Chuntharapai(5),
- Dennis F. Michiel(1),
- Joost J. Oppenheim(1) and
- David J. Kelvin(¶)(1)
- From the (1) Laboratory of Molecular Immunoregulation, Biological Response Modifiers Program,
- (2) PRI/DynCorp, Frederick Cancer Research and Development Center, the
- (3) Preclinical Evaluation Laboratory, Biological Response Modifiers Program, and the
- (4) Laboratory of Experimental Immunology, Biological Response Modifiers Program, NCI, National Institutes of Health, Frederick, Maryland 21702-1201 and
- (5) Genentech, Inc., South San Francisco, California 94080
- § To whom correspondence and reprint requests should be addressed: LMI, BRMP, NCI-FCRDC, Bldg. 560, Rm. 31-19, Frederick, MD 21702-1201. Tel.: 301-846-1347; Fax: 301-846-1673.
Abstract
Two interleukin-8 (IL-8) receptors, α and β, have been identified and cloned. Both receptors are thought to transduce signals by coupling to GTP-binding proteins. The aim of this study is to determine whether the carboxyl terminus (C′) of IL-8 receptor β (IL-8Rβ) is involved in signaling in response to IL-8. We have constructed a number of IL-8Rβ genes that encode truncated forms of the IL-8Rβ. The deletions consisted of amino acids 349-355, 336-355, 325-355, and 317-355 (termed β2, β3, β4, and β5, respectively). 293 human embryonic kidney cells were transfected with the wild type IL-8Rβ (β1) and with these mutants. Cells transfected with the mutated receptors expressed the receptors and bound IL-8 with the same high affinity as cells transfected with the wild type receptor.
The capacity of the mutated receptors to convey functional signals was evaluated by comparing the chemotaxis index of cells expressing the C′-truncated receptors to the index of cells expressing the wild type receptor. The results indicate that while cells expressing β1, β2, β3, and β4 were chemoattracted in response to IL-8, cells expressing β5 did not migrate in response to IL-8 stimulation. Therefore, the data suggest that amino acids 317-324 are involved in signaling by IL-8Rβ.
Footnotes
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↵* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵1 The abbreviations used are:
- IL-8
-
interleukin-8
- IL-8Rα
-
type A receptor for IL-8
- IL-8Rβ
-
type B receptor for IL-8
- N′
-
amino terminus domain of the receptor
- C′
-
carboxyl terminus domain of the receptor
- G protein
-
GTP-binding protein
- BSA
-
bovine serum albumin
- PCR
-
polymerase chain reaction
- WT
-
wild type
- 3i
-
third intracellular
- 7TMR
-
seven transmembrane receptors
- FITC
-
fluorescein isothiocyanate.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
