Preparation of a stable CCL5·CCR5·Gi signaling complex for Cryo-EM analysis

Isaikina, Polina and Tsai, Ching-Ju and Petrovic, Ivana and Rogowski, Marco and Dürr, Alexandra Meng and Grzesiek, Stephan. (2022) Preparation of a stable CCL5·CCR5·Gi signaling complex for Cryo-EM analysis. In: Biomolecular Interactions Part B, 169. Amsterdam, Netherlands, pp. 115-141.

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Official URL: https://edoc.unibas.ch/88607/

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The numerous chemokines and their cognate G protein-coupled chemokine receptors on the surface of leukocytes form a complex signaling network, which regulates the immune response and also other key physiological processes. Currently only a very limited number of structures of chemokine-chemokine receptor complexes have been solved. More structures are needed for the understanding of their mechanism of action and the rational design of drugs against these highly relevant therapeutic targets. Recently, we have determined the cryo-EM structure of the human wild-type CCR5 chemokine receptor, which is also the HIV-1 coreceptor, in its active conformation bound to the chemokine super-agonist [6P4]CCL5 and the heterotrimeric Gi protein. The structure provides the rationale for the sequence-activity relation of agonist and antagonist CCR5 chemokine ligands. In this chapter, we present a detailed protocol for the preparation of the active agonist chemokine-CCR5-Gi complex for cryo-EM studies including quality controls and caveats. As such the protocol may serve as starting point for structural and biophysical studies of other chemokine-chemokine receptor complexes.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Grzesiek)
UniBasel Contributors:Grzesiek, Stephan and Isaikina, Polina and Petrovic, Ivana and Rogowski, Marco and Meng Dürr, Alexandra
Item Type:Book Section
Book Section Subtype:Book Chapter
Series Name:Methods in Cell Biology
Note:Publication type according to Uni Basel Research Database: Book item
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Last Modified:24 May 2023 01:30
Deposited On:20 Jun 2022 07:16

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