التفاصيل البيبلوغرافية
| العنوان: |
Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7. |
| المؤلفون: |
Jakobs, Barbara D., Spannagel, Lisa, Purvanov, Vladimir, Uetz-von Allmen, Edith, Matti, Christoph, Legler, Daniel F. |
| المصدر: |
International Journal of Molecular Sciences; May2019, Vol. 20 Issue 10, p2597, 1p, 1 Diagram, 5 Graphs |
| مصطلحات موضوعية: |
CHEMOKINE receptors, DENDRITIC cells, T cells, G protein coupled receptors, CRYSTAL structure |
| مستخلص: |
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. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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