title: On the mechanism and significance of ligand-induced internalization of human neutrophil chemokine receptors CXCR1 and CXCR2.
abstract: It is well established that leukocyte chemotactic receptors, a subset of G protein-coupled receptors, undergo endocytosis after stimulation by ligand. However, the significance of this phenomenon to cell motility and other important leukocyte functions induced by chemoattractants has not been clearly defined. Here we show that in primary human neutrophils, the threshold levels of agonist required for endocytosis of the chemotactic receptors CXCR1 and CXCR2 were approximately 10-fold or higher than those needed for maximal chemotactic and calcium flux responses. Moreover, when stimulated by agonists at concentrations that are high enough for chemotaxis but too low for receptor endocytosis, neutrophil CXCR1 and CXCR2 could be reactivated in response to repeated application of the same agonist. Both receptors were excluded from Triton X-100-insoluble lipid rafts, and at high agonist concentrations were rapidly endocytosed by a clathrin/rab5/dynamin-dependent pathway. These data support the conclusion that neutrophil migration in response to CXCR1 or CXCR2 agonists is not dependent on endocytosis of CXCR1 or CXCR2. Rather than being integral to the process of cell migration, receptor endocytosis may be a terminal stop signal when cells reach the focus of inflammation where the chemoattractant concentrations are the highest.
authors.affiliation: Laboratory of Molecular Microbiology, NIAID, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Jeremy J
authors.initials: JJ
authors.lastname: Rose
authors.forename: John F
authors.initials: JF
authors.lastname: Foley
authors.forename: Philip M
authors.initials: PM
authors.lastname: Murphy
authors.forename: Sundararajan
authors.initials: S
authors.lastname: Venkatesan
mesh_terms: D018821:CD18 Antigens / Q000096:biosynthesis; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D002633:Chemotaxis; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism; D003505:Cyclodextrins / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D034281:Dynamins / Q000737:chemistry; D004705:Endocytosis; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation; D016209:Interleukin-8 / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D050356:Lipid Metabolism; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis; D021962:Membrane Microdomains; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009504:Neutrophils / Q000378:metabolism; D017830:Octoxynol / Q000494:pharmacology; D010957:Plasmids / Q000378:metabolism; D011487:Protein Conformation; D021381:Protein Transport; D023062:Receptors, Interleukin-8A / Q000378:metabolism*; D023063:Receptors, Interleukin-8B / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D047392:beta-Cyclodextrins; D020696:rab5 GTP-Binding Proteins / Q000737:chemistry
chemical_list: D018821:CD18 Antigens; D003505:Cyclodextrins; D003902:Detergents; D016209:Interleukin-8; D008024:Ligands; D018950:Lipopolysaccharide Receptors; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D017830:Octoxynol; D002784:Cholesterol; D020696:rab5 GTP-Binding Proteins; D034281:Dynamins; D002118:Calcium