KOPS - The Institutional Repository of the University of Konstanz

Biased Signaling of CCL21 and CCL19 Does Not Rely on N-Terminal Differences, but Markedly on the Chemokine Core Domains and Extracellular Loop 2 of CCR7

Biased Signaling of CCL21 and CCL19 Does Not Rely on N-Terminal Differences, but Markedly on the Chemokine Core Domains and Extracellular Loop 2 of CCR7

Cite This

Files in this item

Checksum: MD5:04a5997e8a0918be9d45dc75848e8aa9

JØRGENSEN, Astrid S., Olav LARSEN, Edith UETZ-VON ALLMEN, Michael LÜCKMANN, Daniel F. LEGLER, Thomas M. FRIMURER, Christopher T. VELDKAMP, Gertrud M. HJORTØ, Mette M. ROSENKILDE, 2019. Biased Signaling of CCL21 and CCL19 Does Not Rely on N-Terminal Differences, but Markedly on the Chemokine Core Domains and Extracellular Loop 2 of CCR7. In: Frontiers in Immunology. 10, 2156. eISSN 1664-3224. Available under: doi: 10.3389/fimmu.2019.02156

@article{Jrgensen2019-09-13Biase-46914, title={Biased Signaling of CCL21 and CCL19 Does Not Rely on N-Terminal Differences, but Markedly on the Chemokine Core Domains and Extracellular Loop 2 of CCR7}, year={2019}, doi={10.3389/fimmu.2019.02156}, volume={10}, journal={Frontiers in Immunology}, author={Jørgensen, Astrid S. and Larsen, Olav and Uetz-von Allmen, Edith and Lückmann, Michael and Legler, Daniel F. and Frimurer, Thomas M. and Veldkamp, Christopher T. and Hjortø, Gertrud M. and Rosenkilde, Mette M.}, note={Article Number: 2156} }

2019-09-17T15:19:33Z Hjortø, Gertrud M. Larsen, Olav Legler, Daniel F. 2019-09-13 Jørgensen, Astrid S. Veldkamp, Christopher T. Frimurer, Thomas M. Chemokine receptors play important roles in the immune system and are linked to several human diseases. Targeting chemokine receptors have so far shown very little success owing to, to some extent, the promiscuity of the immune system and the high degree of biased signaling within it. CCR7 and its two endogenous ligands display biased signaling and here we investigate the differences between the two ligands, CCL21 and CCL19, with respect to their biased activation of CCR7. We use bystander bioluminescence resonance energy transfer (BRET) based signaling assays and Transwell migration assays to determine (A) how swapping of domains between the two ligands affect their signaling patterns and (B) how receptor mutagenesis impacts signaling. Using chimeric ligands we find that the chemokine core domains are central for determining signaling outcome as the lack of β-arrestin-2 recruitment displayed by CCL21 is linked to its core domain and not N-terminus. Through a mutagenesis screen, we identify the extracellular domains of CCR7 to be important for both ligands and show that the two chemokines interact differentially with extracellular loop 2 (ECL-2). By using in silico modeling, we propose a link between ECL-2 interaction and CCR7 signal transduction. Our mutagenesis study also suggests a lysine in the top of TM3, K130<sup>3.26</sup>, to be important for G protein signaling, but not β-arrestin-2 recruitment. Taken together, the bias in CCR7 between CCL19 and CCL21 relies on the chemokine core domains, where interactions with ECL-2 seem particularly important. Moreover, TM3 selectively regulates G protein signaling as found for other chemokine receptors. Hjortø, Gertrud M. Lückmann, Michael Lückmann, Michael Uetz-von Allmen, Edith eng Frimurer, Thomas M. Rosenkilde, Mette M. Larsen, Olav Veldkamp, Christopher T. Legler, Daniel F. Jørgensen, Astrid S. Uetz-von Allmen, Edith terms-of-use Rosenkilde, Mette M. Biased Signaling of CCL21 and CCL19 Does Not Rely on N-Terminal Differences, but Markedly on the Chemokine Core Domains and Extracellular Loop 2 of CCR7 2019-09-17T15:19:33Z

Downloads since Sep 17, 2019 (Information about access statistics)

Jorgensen_2-14a6gxbyb03e89.pdf 142

This item appears in the following Collection(s)

Search KOPS


Browse

My Account