Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse
Signaling by the human C-type lectin-like receptor, natural killer (NK) cell inhibitory receptor NKR-P1, has a critical role in many immune-related diseases and cancer. C-type lectin-like receptors have weak affinities to their ligands; therefore, setting up a comprehensive model of NKR-P1-LLT1 inte...
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Science::Biological sciences Membrane Antigen Cell Surface Receptor |
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Science::Biological sciences Membrane Antigen Cell Surface Receptor Bláha, Jan Skálová, Tereza Kalousková, Barbora Skořepa, Ondřej Cmunt, Denis Grobárová, Valéria Pazicky, Samuel Poláchová, Edita Abreu, Celeste Stránský, Jan Kovaľ, Tomáš Dušková, Jarmila Zhao, Yuguang Harlos, Karl Hašek, Jindřich Dohnálek, Jan Vaněk, Ondřej Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse |
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Signaling by the human C-type lectin-like receptor, natural killer (NK) cell inhibitory receptor NKR-P1, has a critical role in many immune-related diseases and cancer. C-type lectin-like receptors have weak affinities to their ligands; therefore, setting up a comprehensive model of NKR-P1-LLT1 interactions that considers the natural state of the receptor on the cell surface is necessary to understand its functions. Here we report the crystal structures of the NKR-P1 and NKR-P1:LLT1 complexes, which provides evidence that NKR-P1 forms homodimers in an unexpected arrangement to enable LLT1 binding in two modes, bridging two LLT1 molecules. These interaction clusters are suggestive of an inhibitory immune synapse. By observing the formation of these clusters in solution using SEC-SAXS analysis, by dSTORM super-resolution microscopy on the cell surface, and by following their role in receptor signaling with freshly isolated NK cells, we show that only the ligation of both LLT1 binding interfaces leads to effective NKR-P1 inhibitory signaling. In summary, our findings collectively support a model of NKR-P1:LLT1 clustering, which allows the interacting proteins to overcome weak ligand-receptor affinity and to trigger signal transduction upon cellular contact in the immune synapse. |
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School of Biological Sciences |
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School of Biological Sciences Bláha, Jan Skálová, Tereza Kalousková, Barbora Skořepa, Ondřej Cmunt, Denis Grobárová, Valéria Pazicky, Samuel Poláchová, Edita Abreu, Celeste Stránský, Jan Kovaľ, Tomáš Dušková, Jarmila Zhao, Yuguang Harlos, Karl Hašek, Jindřich Dohnálek, Jan Vaněk, Ondřej |
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Article |
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Bláha, Jan Skálová, Tereza Kalousková, Barbora Skořepa, Ondřej Cmunt, Denis Grobárová, Valéria Pazicky, Samuel Poláchová, Edita Abreu, Celeste Stránský, Jan Kovaľ, Tomáš Dušková, Jarmila Zhao, Yuguang Harlos, Karl Hašek, Jindřich Dohnálek, Jan Vaněk, Ondřej |
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Bláha, Jan |
title |
Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse |
title_short |
Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse |
title_full |
Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse |
title_fullStr |
Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse |
title_full_unstemmed |
Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse |
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structure of the human nk cell nkr-p1:llt1 receptor:ligand complex reveals clustering in the immune synapse |
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2023 |
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https://hdl.handle.net/10356/171170 |
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sg-ntu-dr.10356-1711702023-10-23T15:32:02Z Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse Bláha, Jan Skálová, Tereza Kalousková, Barbora Skořepa, Ondřej Cmunt, Denis Grobárová, Valéria Pazicky, Samuel Poláchová, Edita Abreu, Celeste Stránský, Jan Kovaľ, Tomáš Dušková, Jarmila Zhao, Yuguang Harlos, Karl Hašek, Jindřich Dohnálek, Jan Vaněk, Ondřej School of Biological Sciences Science::Biological sciences Membrane Antigen Cell Surface Receptor Signaling by the human C-type lectin-like receptor, natural killer (NK) cell inhibitory receptor NKR-P1, has a critical role in many immune-related diseases and cancer. C-type lectin-like receptors have weak affinities to their ligands; therefore, setting up a comprehensive model of NKR-P1-LLT1 interactions that considers the natural state of the receptor on the cell surface is necessary to understand its functions. Here we report the crystal structures of the NKR-P1 and NKR-P1:LLT1 complexes, which provides evidence that NKR-P1 forms homodimers in an unexpected arrangement to enable LLT1 binding in two modes, bridging two LLT1 molecules. These interaction clusters are suggestive of an inhibitory immune synapse. By observing the formation of these clusters in solution using SEC-SAXS analysis, by dSTORM super-resolution microscopy on the cell surface, and by following their role in receptor signaling with freshly isolated NK cells, we show that only the ligation of both LLT1 binding interfaces leads to effective NKR-P1 inhibitory signaling. In summary, our findings collectively support a model of NKR-P1:LLT1 clustering, which allows the interacting proteins to overcome weak ligand-receptor affinity and to trigger signal transduction upon cellular contact in the immune synapse. Published version This study was supported by the Czech Science Foundation grants 15- 15181 S and 18-10687 S to O.V., the Ministry of Education, Youth and Sports of the Czech Republic grant LTC17065 to O.V. (in the frame of the COST Action CA15126 MOBIEU), the Charles University Grant Agency projects 161216 and 1378219 to J.B. and B.K, and the European Regional Development Fund (CZ.02.1.01/0.0/0.0/15_003/0000447). Microscopy was performed in the Laboratory of Confocal and Fluorescence Microscopy, co-financed by the European Regional Development Fund and the state budget of the Czech Republic (projects no. CZ.1.05/4.1.00/ 16.0347 and CZ.2.16/3.1.00/21515) and supported by the CzechBioImaging large RI project LM2018129. Computational resources were supplied by the project “e-Infrastruktura CZ” (e-INFRA LM2018140) provided within the program Projects of Large Research, Development, and Innovations Infrastructures. 2023-10-23T07:12:34Z 2023-10-23T07:12:34Z 2022 Journal Article Bláha, J., Skálová, T., Kalousková, B., Skořepa, O., Cmunt, D., Grobárová, V., Pazicky, S., Poláchová, E., Abreu, C., Stránský, J., Kovaľ, T., Dušková, J., Zhao, Y., Harlos, K., Hašek, J., Dohnálek, J. & Vaněk, O. (2022). Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse. Nature Communications, 13(1), 5022-. https://dx.doi.org/10.1038/s41467-022-32577-6 2041-1723 https://hdl.handle.net/10356/171170 10.1038/s41467-022-32577-6 36028489 2-s2.0-85137120895 1 13 5022 en Nature Communications © The Author(s) 2022.This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/ licenses/by/4.0/. application/pdf |