Protocol

Chemotaxis

Volume 571 of the series Methods in Molecular Biology™ pp 179-198

Date:

Chemokine Receptor Dimerization and Chemotaxis

  • José Miguel Rodríguez-FradeAffiliated withDepartment of Immunology and Oncology, Centro Nacional de Biotecnología
  • , Laura Martinez MuñozAffiliated withDepartment of Immunology and Oncology, Centro Nacional de Biotecnología
  • , Borja L. HolgadoAffiliated withDepartment of Immunology and Oncology, Centro Nacional de Biotecnología
  • , Mario MelladoAffiliated withDepartment of Immunology and Oncology, Centro Nacional de Biotecnología

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Summary

A broad array of biological responses ranging from cell polarization, movement, immune and inflammatory responses, as well as prevention of HIV-1 infection, are triggered by the chemokines, a family of structurally related chemoattractant proteins that bind to specific seven-transmembrane receptors linked to G proteins. Although it was initially believed that chemokine receptors act as monomeric entities, it has now been shown that they function as oligomers. Chemokine receptor homo– and heterodimers are found on the cell membrane; binding to their ligands stabilizes specific receptor conformations and activates distinct signaling cascades. Thorough analysis of the conformations adopted by the receptors at the membrane is therefore a prerequisite for understanding the function of these inflammatory mediators.

For study of the chemokine receptor conformations at the cell surface, we focus here on conventional biochemical and genetic methods, as well as on new imaging techniques such as those based on resonance energy transfer; we also evaluate in vitro and in vivo methods to determine certain chemokine receptor functions.

Key words:

Chemokine Chemokine receptor GPCR Dimerization BRET FRET Chemotaxis