Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. Using X-ray diffraction, the first experimental structures of human LLT1 were determined. Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. The dimeric form follows the classical dimerization mode of human CD69. The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. The hexamer of glycosylated LLT1 consists of three classical dimers. The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form.

Department of Biochemistry Faculty of Science Charles University Prague Hlavova 8 128 40 Praha Czech Republic

Division of Structural Biology The Wellcome Trust Centre for Human Genetics University of Oxford Roosevelt Drive Oxford OX3 7BN England

Institute of Biotechnology Academy of Sciences of the Czech Republic v v i Vídeňská 1083 142 20 Praha 4 Czech Republic

Institute of Macromolecular Chemistry Academy of Sciences of the Czech Republic v v i Heyrovského nám 2 162 06 Praha 6 Czech Republic

See more in PubMed

Aldemir, H., Prod’homme, V., Dumaurier, M.-J., Retiere, C., Poupon, G., Cazareth, J., Bihl, F. & Braud, V. M. (2005). J. Immunol. 175, 7791–7795. PubMed

Aust, J. G., Gays, F., Mickiewicz, K. M., Buchanan, E. & Brooks, C. G. (2009). J. Immunol. 183, 106–116. PubMed

Baker, N. A., Sept, D., Joseph, S., Holst, M. J. & McCammon, J. A. (2001). Proc. Natl Acad. Sci. USA, 98, 10037–10041. PubMed PMC

Bartel, Y., Bauer, B. & Steinle, A. (2013). Front. Immunol. 4, 362. PubMed PMC

Battye, T. G. G., Kontogiannis, L., Johnson, O., Powell, H. R. & Leslie, A. G. W. (2011). Acta Cryst. D67, 271–281. PubMed PMC

Biasini, M., Bienert, S., Waterhouse, A., Arnold, K., Studer, G., Schmidt, T., Kiefer, F., Cassarino, T. G., Bertoni, M., Bordoli, L. & Schwede, T. (2014). Nucleic Acids Res. 42, W252–W258. PubMed PMC

Bláha, J., Pachl, P., Novák, P. & Vaněk, O. (2015). Protein Expres. Purif. 109, 7–13. PubMed

Caligiuri, M. A. (2008). Blood, 112, 461–469. PubMed PMC

Chen, V. B., Arendall, W. B., Headd, J. J., Keedy, D. A., Immormino, R. M., Kapral, G. J., Murray, L. W., Richardson, J. S. & Richardson, D. C. (2010). Acta Cryst. D66, 12–21. PubMed PMC

Chovancova, E., Pavelka, A., Benes, P., Strnad, O., Brezovsky, J., Kozlikova, B., Gora, A., Sustr, V., Klvana, M., Medek, P., Biedermannova, L., Sochor, J. & Damborsky, J. (2012). PLoS Comput. Biol. 8, e1002708. PubMed PMC

Diederichs, K. & Karplus, P. A. (1997). Nature Struct. Mol. Biol. 4, 269–275. PubMed

Dolinsky, T. J., Czodrowski, P., Li, H., Nielsen, J. E., Jensen, J. H., Klebe, G. & Baker, N. A. (2007). Nucleic Acids Res. 35, W522–W525. PubMed PMC

Durocher, Y., Perret, S. & Kamen, A. (2002). Nucleic Acids Res. 30, E9. PubMed PMC

Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. (2010). Acta Cryst. D66, 486–501. PubMed PMC

Evans, P. R. & Murshudov, G. N. (2013). Acta Cryst. D69, 1204–1214. PubMed PMC

Fine, J. H., Chen, P., Mesci, A., Allan, D. S., Gasser, S., Raulet, D. H. & Carlyle, J. R. (2010). Cancer Res. 70, 7102–7113. PubMed PMC

French, S. & Wilson, K. (1978). Acta Cryst. A34, 517–525.

Germain, C., Bihl, F., Zahn, S., Poupon, G., Dumaurier, M. J., Rampanarivo, H. H., Padkjaer, S. B., Spee, P. & Braud, V. M. (2010). J. Biol. Chem. 285, 36207–36215. PubMed PMC

Germain, C., Meier, A., Jensen, T., Knapnougel, P., Poupon, G., Lazzari, A., Neisig, A., Håkansson, K., Dong, T., Wagtmann, N., Galsgaard, E. D., Spee, P. & Braud, V. M. (2011). J. Biol. Chem. 286, 37964–37975. PubMed PMC

Gouet, P., Robert, X. & Courcelle, E. (2003). Nucleic Acids Res. 31, 3320–3323. PubMed PMC

Grobárová, V., Benson, V., Rozbeský, D., Novák, P. & Černý, J. (2013). Immunol. Lett. 156, 110–117. PubMed

Grueninger-Leitch, F., D’Arcy, A., D’Arcy, B. & Chène, C. (1996). Protein Sci. 5, 2617–2622. PubMed PMC

Huarte, E., Cubillos-Ruiz, J. R., Nesbeth, Y. C., Scarlett, U. K., Martinez, D. G., Engle, X. A., Rigby, W. F., Pioli, P. A., Guyre, P. M. & Conejo-Garcia, J. R. (2008). Blood, 112, 1259–1268. PubMed PMC

Kabsch, W. & Sander, C. (1983). Biopolymers, 22, 2577–2637. PubMed

Kamishikiryo, J., Fukuhara, H., Okabe, Y., Kuroki, K. & Maenaka, K. (2011). J. Biol. Chem. 286, 23823–23830. PubMed PMC

Keegan, R. M., Long, F., Fazio, V. J., Winn, M. D., Murshudov, G. N. & Vagin, A. A. (2011). Acta Cryst. D67, 313–323. PubMed PMC

Kolenko, P., Rozbeský, D., Vaněk, O., Bezouška, K., Hašek, J. & Dohnálek, J. (2011). Acta Cryst. F67, 1519–1523. PubMed PMC

Kolenko, P., Rozbeský, D., Vaněk, O., Kopecký, V. Jr, Hofbauerová, K., Novák, P., Pompach, P., Hašek, J., Skálová, T., Bezouška, K. & Dohnálek, J. (2011). J. Struct. Biol. 175, 434–441. PubMed

Kolenko, P., Skálová, T., Vaněk, O., Štěpánková, A., Dušková, J., Hašek, J., Bezouška, K. & Dohnálek, J. (2009). Acta Cryst. F65, 1258–1260. PubMed PMC

Krissinel, E. & Henrick, K. (2007). J. Mol. Biol. 372, 774–797. PubMed

Kveberg, L., Dai, K.-Z., Westgaard, I. H., Daws, M. R., Fossum, S., Naper, C. & Vaage, J. T. (2009). Eur. J. Immunol. 39, 541–551. PubMed

Lanier, L. L. (2008). Nature Immunol. 9, 495–502. PubMed PMC

Lebedev, A. A. & Isupov, M. N. (2014). Acta Cryst. D70, 2430–2443. PubMed

Leslie, A. G. W. & Powell, H. R. (2007). Evolving Methods for Macromolecular Crystallography, edited by R. J. Read & J. L. Sussman, pp. 41–51. Dordrecht: Springer.

Li, H., Robertson, A. D. & Jensen, J. H. (2005). Proteins, 61, 704–721. PubMed

Li, Y., Wang, Q., Chen, S., Brown, P. H. & Mariuzza, R. A. (2013). Proc. Natl Acad. Sci. USA, 110, 11505–11510. PubMed PMC

Llera, A. S., Viedma, F., Sánchez-Madrid, F. & Tormo, J. (2001). J. Biol. Chem. 276, 7312–7319. PubMed

Long, F., Vagin, A. A., Young, P. & Murshudov, G. N. (2008). Acta Cryst. D64, 125–132. PubMed PMC

Mueller, U., Darowski, N., Fuchs, M. R., Förster, R., Hellmig, M., Paithankar, K. S., Pühringer, S., Steffien, M., Zocher, G. & Weiss, M. S. (2012). J. Synchrotron Rad. 19, 442–449. PubMed PMC

Murshudov, G. N., Skubák, P., Lebedev, A. A., Pannu, N. S., Steiner, R. A., Nicholls, R. A., Winn, M. D., Long, F. & Vagin, A. A. (2011). Acta Cryst. D67, 355–367. PubMed PMC

Natarajan, K., Sawicki, M. W., Margulies, D. H. & Mariuzza, R. A. (2000). Biochemistry, 39, 14779–14786. PubMed

Ortega, A., Amorós, D. & García de la Torre, J. (2011). Biophys. J. 101, 892–898. PubMed PMC

Otwinowski, Z. & Minor, W. (1997). Methods Enzymol. 276, 307–326. PubMed

Reeves, P. J., Callewaert, N., Contreras, R. & Khorana, H. G. (2002). Proc. Natl Acad. Sci. USA, 99, 13419–13424. PubMed PMC

Rosen, D. B., Bettadapura, J., Alsharifi, M., Mathew, P. A., Warren, H. S. & Lanier, L. L. (2005). J. Immunol. 175, 7796–7799. PubMed

Rosen, D. B., Cao, W., Avery, D. T., Tangye, S. G., Liu, Y.-J., Houchins, J. P. & Lanier, L. L. (2008). J. Immunol. 180, 6508–6517. PubMed PMC

Roth, P., Mittelbronn, M., Wick, W., Meyermann, R., Tatagiba, M. & Weller, M. (2007). Cancer Res. 67, 3540–3544. PubMed

Satkunanathan, S., Kumar, N., Bajorek, M., Purbhoo, M. A., Culley, F. J. & Lyles, D. (2014). J. Virol. 88, 2366–2373. PubMed PMC

Schuck, P. (2003). Anal. Biochem. 320, 104–124. PubMed

Sievers, F., Wilm, A., Dineen, D., Gibson, T. J., Karplus, K., Li, W., Lopez, R., McWilliam, H., Remmert, M., Söding, J., Thompson, J. D. & Higgins, D. G. (2011). Mol. Syst. Biol. 7, 539. PubMed PMC

Skálová, T., Kotýnková, K., Dušková, J., Hašek, J., Koval, T., Kolenko, P., Novák, P., Man, P., Hanč, P., Vaněk, O., Bezouška, K. & Dohnálek, J. (2012). J. Immunol. 189, 4881–4889. PubMed

Sovová, Ž., Kopecký, V. Jr, Pazderka, T., Hofbauerová, K., Rozbeský, D., Vaněk, O., Bezouška, K. & Ettrich, R. (2011). J. Mol. Model. 17, 1353–1370. PubMed

Strong, M., Sawaya, M. R., Wang, S., Phillips, M., Cascio, D. & Eisenberg, D. (2006). Proc. Natl Acad. Sci. USA, 103, 8060–8065. PubMed PMC

Sullivan, L. C., Clements, C. S., Beddoe, T., Johnson, D., Hoare, H. L., Lin, J., Huyton, T., Hopkins, E. J., Reid, H. H., Wilce, M. C. J., Kabat, J., Borrego, F., Coligan, J. E., Rossjohn, J. & Brooks, A. G. (2007). Immunity, 27, 900–911. PubMed

Thompson, J. D., Higgins, D. G. & Gibson, T. J. (1994). Nucleic Acids Res. 22, 4673–4680. PubMed PMC

Vagin, A. & Teplyakov, A. (2010). Acta Cryst. D66, 22–25. PubMed

Vaněk, O. et al. (2008). FEBS J. 275, 5589–5606. PubMed

Vivier, E., Raulet, D. H., Moretta, A., Caligiuri, M. A., Zitvogel, L., Lanier, L. L., Yokoyama, W. M. & Ugolini, S. (2011). Science, 331, 44–49. PubMed PMC

Vivier, E., Tomasello, E., Baratin, M., Walzer, T. & Ugolini, S. (2008). Nature Immunol. 9, 503–510. PubMed

Voigt, S., Mesci, A., Ettinger, J., Fine, J. H., Chen, P., Chou, W. & Carlyle, J. R. (2007). Immunity, 26, 617–627. PubMed

Weiss, M. S. (2001). J. Appl. Cryst. 34, 130–135.

Williams, K. J., Wilson, E., Davidson, C. L., Aguilar, O. A., Fu, L., Carlyle, J. R. & Burshtyn, D. N. (2012). J. Immunol. 188, 4980–4991. PubMed

Winn, M. D. et al. (2011). Acta Cryst. D67, 235–242. PubMed

Yokoyama, W. M. & Plougastel, B. F. M. (2003). Nature Rev. Immunol. 3, 304–316. PubMed

Zelensky, A. N. & Gready, J. E. (2005). FEBS J. 272, 6179–6217. PubMed

Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse

Protein purification strategies must consider downstream applications and individual biological characteristics

Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells

Production of recombinant soluble dimeric C-type lectin-like receptors of rat natural killer cells