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The ligand binding site of the formyl peptide receptor maps in the transmembrane region.

H M Miettinen, J S Mills, J M Gripentrog, E A Dratz, B L Granger and A J Jesaitis
J Immunol October 15, 1997, 159 (8) 4045-4054;
H M Miettinen
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J S Mills
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J M Gripentrog
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E A Dratz
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B L Granger
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A J Jesaitis
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Abstract

We propose that the N-formyl-Met-Leu-Phe binding site in the human neutrophil formyl peptide receptor (FPR) lies in the predicted transmembrane region. We examined the expression, binding, and G protein coupling of 28 mutated forms of FPR in stably transfected Chinese hamster ovary cells. The amino acids we mutated are: 1) predicted to be oriented toward the interhelical space; 2) analogous to those required for ligand binding in various other G protein-coupled receptors; 3) divergent from lipoxin A4 receptor, a low affinity receptor for formylated peptides; and 4) either highly conserved or divergent in other G protein-coupled receptors. Some mutations resulted in intracellular retention, suggesting that the receptors were misfolded. Most mutated receptors that were transported to the plasmalemma bound f-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein with affinities similar to the wild-type receptor (Kd = 6 nM). However, mutations L78A (helix II), D106N, L109A (helix III), T157A (helix IV), R201A, I204Y, and R205A (helix V), W254A and Y257A (helix VI), and F291A (helix VII) resulted in reduced affinities (Kd = 30-128 nM). Of these mutations, D106N, R201A, and R205A also appeared to affect G protein coupling, suggesting that these residues may also be involved in signal transduction and/or are essential for proper folding of the molecule. Some of the FPR residues that appeared to be involved in binding of formylated peptides were located at sites analogous to those identified in ligand binding to certain other G protein-coupled receptors. It is thus possible that several G protein-coupled receptors have a common placement of ligand-binding amino acids.

  • Copyright © 1997 by American Association of Immunologists

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The Journal of Immunology
Vol. 159, Issue 8
15 Oct 1997
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The ligand binding site of the formyl peptide receptor maps in the transmembrane region.
H M Miettinen, J S Mills, J M Gripentrog, E A Dratz, B L Granger, A J Jesaitis
The Journal of Immunology October 15, 1997, 159 (8) 4045-4054;

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The ligand binding site of the formyl peptide receptor maps in the transmembrane region.
H M Miettinen, J S Mills, J M Gripentrog, E A Dratz, B L Granger, A J Jesaitis
The Journal of Immunology October 15, 1997, 159 (8) 4045-4054;
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Print ISSN 0022-1767        Online ISSN 1550-6606