title: CXC chemokine receptor 4 signaling upon co-activation with stromal cell-derived factor-1α and ubiquitin.
abstract: Recently, we reported that extracellular ubiquitin functions as another agonist of CXC chemokine receptor (CXCR)4. Whereas the cognate CXCR4 ligand, stromal cell-derived factor (SDF)-1α, is also a CXCR7 agonist, ubiquitin does not bind to CXCR7. Because both ligands are present in the extracellular environment, co-activation of CXCR4 appears to be physiologically relevant. CXCR4 mediated effects of ubiquitin, however, are not well understood and consequences of co-activation of CXCR4 with both ligands are unknown. Utilizing proximity ligation assays and flow cytometry, we detected CXCR4, but not CXCR7, on the cell surface of THP-1 cells, which suggests that confounding effects of CXCR7 are unlikely. Time course and magnitude of reduction of cell surface CXCR4 expression were comparable after stimulation of THP-1 cells with both ligands. SDF-1α was more efficacious than ubiquitin to mobilize Ca(2+). Co-stimulation of THP-1 cells with both ligands resulted in synergistic effects on Ca(2+) fluxes at suboptimal ligand concentrations. Homologous desensitization of Ca(2+) fluxes was detectable with both ligands. SDF-1α pre-stimulation desensitized ubiquitin induced Ca(2+) fluxes, but not vice versa. Effects of SDF-1α and ubiquitin on cAMP levels, Akt and ERK1/2 phosphorylation and chemotactic responses were additive. The chemotactic activities of ubiquitin and SDF-1α were sensitive to AMD3100, pertussis toxin, U73122, LY94002 and U0126. These data suggest that CXCR4 activation with SDF-1α and ubiquitin results in partially synergistic effects on cellular signaling events and in differential effects on receptor desensitization. The ligand ratio that is present in the extracellular environment may contribute to the regulation of CXCR4 mediated functions.
authors.affiliation: Department of Surgery, Loyola University Chicago, Stritch School of Medicine, United States.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Tripathi
authors.affiliation: Department of Surgery, Loyola University Chicago, Stritch School of Medicine, United States.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Davis
authors.affiliation: Department of Surgery, Loyola University Chicago, Stritch School of Medicine, United States.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Staren
authors.affiliation: Department of Biochemistry, Medical College of Wisconsin, United States.
authors.forename: Brian F
authors.initials: BF
authors.lastname: Volkman
authors.affiliation: Department of Surgery, Loyola University Chicago, Stritch School of Medicine, United States; Department of Molecular Pharmacology & Therapeutics, Loyola University Chicago, Stritch School of Medicine, United States. Electronic address: mmajetschak@lumc.edu.
authors.forename: Matthias
authors.initials: M
authors.lastname: Majetschak
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D054377:Chemokine CXCL12 / Q000494:pharmacology*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D054387:Receptors, CXCR / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D025801:Ubiquitin / Q000494:pharmacology*
chemical_list: C501896:ACKR3 protein, human; C501898:Ackr3 protein, rat; C527276:CXCR4 protein, human; D054377:Chemokine CXCL12; C486264:Cxcr4 protein, rat; D054387:Receptors, CXCR; D019718:Receptors, CXCR4; D025801:Ubiquitin; D002118:Calcium
keywords: CXCL12; CXCR7; Calcium; Chemotaxis; Cyclic AMP