title: Imaging chemokine receptor dimerization with firefly luciferase complementation.
abstract: Seven-transmembrane (G-protein coupled) receptors are key regulators of normal physiology and a large number of diseases, and this family of receptors is the target for almost half of all drugs. Cell culture models suggest that homodimerization and heterodimerization of 7-transmembrane receptors regulate processes including specificity of ligand binding and activation of downstream signaling pathways, making receptor dimerization a critical determinant of receptor biology and a promising new therapeutic target. To monitor receptor dimerization in cell-based assays and living animals, we developed a protein fragment complementation assay based on firefly luciferase to investigate dimerization of chemokine receptors CXCR4 and CXCR7, two 7-transmembrane receptors with central functions in normal development, cancer, and other diseases. Treatment with chemokine ligands and pharmacologic agents produced time- and dose-dependent changes in reporter signal. Chemokines regulated reporter bioluminescence for CXCR4 or CXCR7 homodimers without affecting signals from receptor heterodimers. In a tumor xenograft model of breast cancer, we used bioluminescence imaging to measure changes in receptor homodimerization in response to pharmacologic agents. This technology should be valuable for analyzing function and therapeutic modulation of receptor dimerization in intact cells and living mice.
authors.affiliation: Center for Molecular Imaging, Department of Radiology, University of Michigan Medical School, 109 Zina Pitcher Pl., A526 BSRB, Ann Arbor, MI 48109-2200, USA. gluker@umich.edu
authors.forename: Kathryn E
authors.initials: KE
authors.lastname: Luker
authors.forename: Mudit
authors.initials: M
authors.lastname: Gupta
authors.forename: Gary D
authors.initials: GD
authors.lastname: Luker
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism; D002460:Cell Line; D018925:Chemokines / Q000378:metabolism; D005260:Female; D005453:Fluorescence; D005816:Genetic Complementation Test / Q000379:methods; D006801:Humans; D049409:Luciferases, Firefly / Q000378:metabolism*; D049408:Luminescent Agents / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D054387:Receptors, CXCR / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000378:metabolism*
chemical_list: C501896:ACKR3 protein, human; C527276:CXCR4 protein, human; D018925:Chemokines; D049408:Luminescent Agents; D054387:Receptors, CXCR; D019718:Receptors, CXCR4; D049409:Luciferases, Firefly