Chemokine receptor signaling

Physiological Roles of Chemokine Receptor Signaling in Nervous System... 

Ottoson NC, Pribila JT, Chan AS, Shimizu Y. Cutting edge T cell migration regulated by CXCR4 chemokine receptor signaling to ZAP-70 tyrosine kinase. J Immunol 2001 167(4) 1857-1861. 

Significance of HIV-1 gpl20-Induced Chemokine Receptor Signaling... 

Del Como M, Liu QH, Schols D, et al. HIV-1 gpl20 and chemokine activation of Pyk2 and mitogen-activated protein kinases in primary macrophages mediated by calcium-dependent, pertussis toxin-insensitive chemokine receptor signaling. Blood 2001 98(10) 2909—2916. 

There has been a great deal of interest in chemokine receptor signalling in cancer in terms of the specific pathways and effector molecules that regulate cell survival, proliferation, chemotaxis, migration and adhesion. It is clear that a large number of downstream effector molecules that are regulated by chemo-kines may account for the multiple effects of these receptors in the pathobiology... 

Many aspects of G-protein-mediated signalling remain to be elucidated, and to gain a broader understanding of the functional roles of chemokine receptor signalling pathways in human breast and potentially other cancer cells,... 

This chapter will focus on how chemokine receptors signal and what role they play in immunoregulation, with an aim toward providing a foundation of basic concepts for the other chapters in this volume, dealing with specific roles of chemokines and chemokine receptors in lung disease. Because of space constraints, the literature cited will be restricted to more recent work and seminal earlier studies. Additional references can be found in a number of excellent reviews (2,4,5). 

Chemokine Receptors Signal Primarily Through Gi-Linked Pathways... 

We describe the overall approach for generating chck beetle luciferase complementation reporters for live-cell bioluminescence imaging of chemokine receptor signaling. We refer readers to other standard texts, such as Methods in Molecular Biology, for detailed protocols for molecular biology procedures including PCR and ligations. 

In this chapter, we describe protocols to set up BRET-based arrestin recruitment test systems. We provide suggestions for troubleshooting and identify experimental parameters that can be varied. We also discuss potential pitfalls and, importandy, some limitations of BRET as a quantitatively comparative method. Indeed, BRET is now widely used as a proximity-based assay to detect arrestin recruitment, but it is often overlooked that, besides proximity, the relative orientations of the probes crucially determines BRET efficiency. This somewhat complicates the accurate interpretation of BRET data, but careful setup and combination of different BRET systems hold promise for the study of yet underexplored questions in chemokine receptor signaling. 

Figure 3 Chemokine receptor signaling pathways. Rough overview of the signaling pathways mediated by chemokine receptors that lead to migration or contribute to cell survival and proliferation. For outcomes highlighted in pink, assays are described in is an artificial G protein used in an iPs turnover assay to mea-...

In the foUowing, we wiU describe assays to assess several outcomes of chemokine receptor signaling, starting with G protein activation, secondary messenger production, and P-arrestin recruitment, continuing with further downstream signaUng (e.g., ERK phosphorylation), and finaUy assessing the chemotactic response. 

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