Type of Publication: | Journal article |
Publication status: | Published |
URI (citable link): | http://nbn-resolving.de/urn:nbn:de:bsz:352-2-13zy0347kj6w93 |
Author: | Jakobs, Barbara D.; Spannagel, Lisa; Purvanov, Vladimir; Uetz-von Allmen, Edith; Matti, Christoph; Legler, Daniel F. |
Year of publication: | 2019 |
Published in: | International Journal of Molecular Sciences ; 20 (2019), 10. - 2597. - eISSN 1422-0067 |
Pubmed ID: | 31137829 |
DOI (citable link): | https://dx.doi.org/10.3390/ijms20102597 |
Summary: |
The chemokine receptor CCR7 plays a pivotal role in health and disease. In particular, CCR7 controls homing of antigen-bearing dendritic cells and T cells to lymph nodes, where adaptive immune responses are initiated. However, CCR7 also guides T cells to inflamed synovium and thereby contributes to rheumatoid arthritis and promotes cancer cell migration and metastasis formation. Nanobodies have recently emerged as versatile tools to study G-protein-coupled receptor functions and are being tested in diagnostics and therapeutics. In this study, we designed a strategy to engineer novel nanobodies recognizing human CCR7. We generated a nanobody library based on a solved crystal structure of the nanobody Nb80 recognizing the β2-adrenergic receptor (β2AR) and by specifically randomizing two segments within complementarity determining region 1 (CDR1) and CDR3 of Nb80 known to interact with β2AR. We fused the nanobody library to one half of split-YFP in order to identify individual nanobody clones interacting with CCR7 fused to the other half of split-YFP using bimolecular fluorescence complementation. We present three novel nanobodies, termed Nb1, Nb5, and Nb38, that recognize human CCR7 without interfering with G-protein-coupling and downstream signaling. Moreover, we were able to follow CCR7 trafficking upon CCL19 triggering using Nb1, Nb5, and Nb38.
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Subject (DDC): | 570 Biosciences, Biology |
Keywords: | chemokine receptor; CCR7; chemokines; CCL19; nanobodies; 2-adrenergic receptor; bimolecular fluorescence complementation; split-luciferase complementation |
Link to License: | In Copyright |
Bibliography of Konstanz: | Yes |
Refereed: | Yes |
JAKOBS, Barbara D., Lisa SPANNAGEL, Vladimir PURVANOV, Edith UETZ-VON ALLMEN, Christoph MATTI, Daniel F. LEGLER, 2019. Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7. In: International Journal of Molecular Sciences. 20(10), 2597. eISSN 1422-0067. Available under: doi: 10.3390/ijms20102597
@article{Jakobs2019-05-27Engin-46455, title={Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7}, year={2019}, doi={10.3390/ijms20102597}, number={10}, volume={20}, journal={International Journal of Molecular Sciences}, author={Jakobs, Barbara D. and Spannagel, Lisa and Purvanov, Vladimir and Uetz-von Allmen, Edith and Matti, Christoph and Legler, Daniel F.}, note={Article Number: 2597} }
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