Identification of Putative Sites of Interaction between the Human Formyl Peptide Receptor and G Protein

Miettinen, Heini M.; Gripentrog, Jeannie M.; Mason, Meta M.; Jesaitis, Algirdas J.

doi:10.1074/jbc.274.39.27934

Cited by 36 publications

(40 citation statements)

References 55 publications

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“…Although selective regions of IL2 and IL3 are essential for G protein recognition, the cooperative interaction of these domains has also been shown to be critical for conferring coupling specificity (41)(42)(43)(44). Our finding that the combined, but not individual, replacement of IL2 and IL3 motifs is required to switch SSTR2 coupling to NHE1 supports the requirement for a cooperative interaction between these two domains.…”

Section: Discussionsupporting

confidence: 68%

Conserved Motifs in Somatostatin, D2-dopamine, and α2B-Adrenergic Receptors for Inhibiting the Na-H Exchanger, NHE1

Lin

Varma

Joubel

et al. 2003

Journal of Biological Chemistry

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Receptor subtypes within families of G proteincoupled receptors that are activated by similar ligands can regulate distinct intracellular effectors. We identified conserved motifs within intracellular domains 2 and 3 of selective subtypes of several G protein-coupled receptor families that confer coupling to the Na-H exchanger, NHE1. A T(s,p)V motif within intracellular domain 2 and a QQ(r) motif within intracellular domain 3 are shared by the somatostatin receptor subtypes SSTR1, -3, and -4, which couple to the inhibition of NHE1, but not by SSTR2 and -5, which do not signal to NHE1. Only the collective substitution of cognate SSTR2 residues with these two motifs conferred the ability of mutant SSTR2 to inhibit NHE1. Both motifs are present in D 2 -dopamine receptors, which inhibit NHE1, and in ␣ 2B -adrenergic receptors, which couple to the inhibition of NHE1, but not in ␣ 2A -adrenergic receptors, which do not regulate NHE1. These findings indicate that motifs shared by different subfamilies of G protein-coupled receptors, but not necessarily by receptor subtypes within a subfamily, can confer coupling to a common effector.Members of the superfamily of G protein-coupled receptors (GPCRs) 1 share a heptahelical transmembrane topology and similar mechanisms for transducing signals through trimeric G proteins to intracellular effectors. Subfamilies of GPCRs, distinguished by sequence identity, generally include several receptor isoforms or subtypes that can be activated by similar ligands. Although receptor subtypes within a GPCR subfamily often regulate similar intracellular effectors, they can also signal to distinct effectors and signaling networks through a process termed receptor subtype-specific signaling. Using the subfamily of somatostatin receptors as a model, the objective of this study was to determine a molecular basis for subtypespecific signaling by GPCRs.The somatostatin receptor family includes five subtypes (SSTR1 to SSTR5) (1-5) that belong to the class A group of GPCRs. Each member is activated by the neuroendocrine peptide somatostatin-14 (SST). SST inhibits diverse cell functions such as hormone secretion, neurotransmitter release, smooth muscle contractility, and cell proliferation. These cellular effects are mediated by coupling of the five SSTR subtypes to similar as well as distinct effectors and signaling networks. All five subtypes couple to the inhibition of adenylyl cyclase (6) and the activation of tyrosine phosphatases (7-13). Different SSTR subtypes, however, also regulate distinct effectors. SSTR2 to -5, but not SSTR1, activate the GIRK1 inwardly rectifying K ϩ channel (14), SSTR1 and -2 inhibit voltage-activated Ca 2ϩ channels (15-17), SSTR2 and -5 stimulate phospholipase C activity (8, 18), and only SSTR4 has been shown to stimulate phospholipase A 2 (19).SST acting at endogenous SSTRs in enteric endocrine cells (20) and hepatic cells (21) also inhibits the activity of the ubiquitously expressed Na-H exchanger NHE1. When heterologously expressed in fibroblasts, SSTR1, but not S...

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Section: Discussionsupporting

confidence: 68%

Conserved Motifs in Somatostatin, D2-dopamine, and α2B-Adrenergic Receptors for Inhibiting the Na-H Exchanger, NHE1

Lin

Varma

Joubel

et al. 2003

Journal of Biological Chemistry

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“…First, the [ 3 H]fMLP binding assay, as other agonist binding assays (25), largely underestimates the actual GPCR expression level (21). Second, high-affinity [ 3 H]fMLP binding depends on intact FPR/G i protein coupling (19,20,26), but we assumed that G i protein coupling is defective in FPR-F110S and FPR-C126W. Fig.…”

Section: Different Electrophoretic Mobility and Glycosylation Of Fpr-mentioning

confidence: 99%

“…A G i protein coupling defect in FPR-C126W was not unexpected since the second intracellular loop is involved in G i protein coupling (18) and since FPR-C126S exhibits a similar functional phenotype as FPR-C126W (Figs. 2-6) (19).…”

Section: Inefficiency Of Fpr-f110s and Fpr-c126w In Stimulating Gtp␥smentioning

confidence: 99%

“…Defective (19,20,26). In previous studies on various GPCRs including the FPR, we already showed that G␣ i2 is expressed at high levels (ϳ200 -450 pmol/mg) in Sf9 membranes (21,22,31).…”

Section: Different Electrophoretic Mobility and Glycosylation Of Fpr-mentioning

confidence: 99%

“…1). The second intracellular loop of the FPR is important for G i protein coupling (18), and a C126S mutation uncouples the FPR from G i proteins (19). Based on all these findings, we developed the hypothesis that the underlying cause of LJP is defective G i protein coupling of the FPR.…”

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confidence: 99%

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Defective Gi Protein Coupling in Two Formyl Peptide Receptor Mutants Associated with Localized Juvenile Periodontitis

Seifert

Wenzel-Seifert

2001

Journal of Biological Chemistry

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Neutrophils play an important role in host defense against bacterial infections and in the pathogenesis of various inflammatory diseases (1, 2). Neutrophils express GPCRs 1 for the chemoattractants fMLP, complement C5a, interleukin-8, leukotriene B 4 , and platelet-activating factor (3-6). Chemoattractant receptors couple to G i proteins to activate phospholipase C␤2 and phosphoinositide 3-kinase-␥, with subsequent stimulation of chemotaxis, oxygen radical formation, and granule release (3, 7-9). Disruption of the FPR gene in mice increases susceptibility to infection by Listeria monocytogenes (10). However, although at cellular and molecular levels, the human FPR is one of the most extensively studied GPCRs (3-6, 8, 11), the in vivo role of the FPR in man has remained elusive.LJP is a defined periodontal disease that begins at ϳ11-15 years of age, affects the permanent incisors and/or first molars, and is associated with the presence of Actinobacillus actinomycetemcomitans in subgingival pockets. LJP is not associated with other diseases, and there is evidence that LJP is caused by a genetic defect (12, 13). It has been known for a long time that neutrophils from LJP patients show reduced binding of the agonist fMLP and reduced fMLP-induced chemotaxis (14 -16). Genetic analysis of 30 LJP patients revealed that 29 of those patients possess a F110S and/or C126W mutation in the FPR gene (17). In contrast, none of 31 patients with adult periodontitis and none of 20 control subjects possess a F110S or C126W mutation (17). The F110S mutation resides in the third transmembrane domain, whereas the C126W mutation resides in the second intracellular loop (Fig. 1). The second intracellular loop of the FPR is important for G i protein coupling (18), and a C126S mutation uncouples the FPR from G i proteins (19). Based on all these findings, we developed the hypothesis that the underlying cause of LJP is defective G i protein coupling of the FPR. To test this hypothesis, we engineered FLAG epitopetagged FPR-F110S and FPR-C126W and analyzed coupling of these FPR mutants and FPR-WT to the G protein G␣ i2 ␤ 1 ␥ 2 using Sf9 insect cells as the expression system. In previous studies, we already showed that Sf9 insect cells are a very sensitive system for studying G i protein coupling of chemoattractant receptors (20 -22). Here we report that FPR-F110S and FPR-C126W show an almost complete defect and a complete defect in G i protein coupling, respectively. The discrete clinical symptoms associated with the defective FPR indicate that the loss of FPR function in host defense is, for the most part, readily compensated. EXPERIMENTAL PROCEDURESMaterials-The baculovirus encoding G␣ i2 was kindly provided by Dr. A. G. Gilman (Department of Pharmacology, University of Southwestern Medical Center, Dallas, TX). Recombinant baculovirus encoding the unmodified versions of the G protein ␤ 1 ␥ 2 subunits was a kind gift of Dr.

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Alanine scanning mutagenesis of CCR3 reveals that the three intracellular loops are essential for functional receptor expression

et al. 2002

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Identification of Putative Sites of Interaction between the Human Formyl Peptide Receptor and G Protein

Abstract: The N-formyl peptide receptor (FPR; also called FPR1) 1 is a seven-transmembrane domain G protein-coupled receptor (GPCR) expressed mainly in myeloid cells (for a review, see Ref.

Cited by 36 publications

References 55 publications

Conserved Motifs in Somatostatin, D2-dopamine, and α2B-Adrenergic Receptors for Inhibiting the Na-H Exchanger, NHE1

Conserved Motifs in Somatostatin, D2-dopamine, and α2B-Adrenergic Receptors for Inhibiting the Na-H Exchanger, NHE1

Defective Gi Protein Coupling in Two Formyl Peptide Receptor Mutants Associated with Localized Juvenile Periodontitis

Alanine scanning mutagenesis of CCR3 reveals that the three intracellular loops are essential for functional receptor expression

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