Crystallography has been the routine technique for studying high-resolution structures of proteins for over five decades. A major bottleneck in structure determination of macromolecules is obtaining crystals of a size and quality suitable for single-crystal X-ray crystallography experiments. Many challenging proteins either fail to grow into crystals or fail to grow into crystals of a size suitable for obtaining high-resolution structures using conventional X-ray crystallography. When it comes to smaller crystals, they can be used either for seeding to get larger crystals or for serial crystallography and electron diffraction for obtaining the structures. For both purposes, a limiting step is to non-invasively image such small crystals of sub-micrometre dimensions and to screen the conditions where such crystals prevail. Here we use cathodoluminescence-based (CL-based) nanoscopy to image protein nanocrystals. We show that crystals of micrometre and submicrometre dimensions can be non-invasively imaged by the CL-based nanoscope. The results presented here demonstrate the feasibility of non-invasive imaging of protein crystals with sub-100 nm resolution.

Extreme Light Infrastructure Za Radnici 835 25241Dolni Brezany Czechia

Research Institute of Electronics Shizuoka University 3 5 1 Johoku Chuo ku Hamamatsu432 8561 Japan

Zobrazit více v PubMed

Beale, E. V., Warren, A. J., Trincão, J., Beilsten-Edmands, J., Crawshaw, A. D., Sutton, G., Stuart, D. & Evans, G. (2020). IUCrJ, 7, 500–508. PubMed PMC

Bischak, C. G., Wai, R. B., Cherqui, C., Busche, J. A., Quillin, S. C., Hetherington, C. L., Wang, Z., Aiello, C. D., Schlom, D. G., Aloni, S., Ogletree, D. F., Masiello, D. J. & Ginsberg, N. S. (2017). ACS Nano, 11, 10583–10590. PubMed

Bodenstaff, E. R., Hoedemaeker, F. J., Kuil, M. E., de Vrind, H. P. M. & Abrahams, J. P. (2002). Acta Cryst. D58, 1901–1906. PubMed

Canfield, R. E. (1963). J. Biol. Chem.238, 2698–2707. PubMed

Chapman, H. N., Fromme, P., Barty, A., White, T. A., Kirian, R. A., Aquila, A., Hunter, M. S., Schulz, J., DePonte, D. P., Weierstall, U., Doak, R. B., Maia, F. R. N. C., Martin, A. V., Schlichting, I., Lomb, L., Coppola, N., Shoeman, R. L., Epp, S. W., Hartmann, R., Rolles, D., Rudenko, A., Foucar, L., Kimmel, N., Weidenspointner, G., Holl, P., Liang, M., Barthelmess, M., Caleman, C., Boutet, S., Bogan, M. J., Krzywinski, J., Bostedt, C., Bajt, S., Gumprecht, L., Rudek, B., Erk, B., Schmidt, C., Hömke, A., Reich, C., Pietschner, D., Strüder, L., Hauser, G., Gorke, H., Ullrich, J., Herrmann, S., Schaller, G., Schopper, F., Soltau, H., Kühnel, K., Messerschmidt, M., Bozek, J. D., Hau-Riege, S. P., Frank, M., Hampton, C. Y., Sierra, R. G., Starodub, D., Williams, G. J., Hajdu, J., Timneanu, N., Seibert, M. M., Andreasson, J., Rocker, A., Jönsson, O., Svenda, M., Stern, S., Nass, K., Andritschke, R., Schröter, C., Krasniqi, F., Bott, M., Schmidt, K. E., Wang, X., Grotjohann, I., Holton, J. M., Barends, T. R. M., Neutze, R., Marchesini, S., Fromme, R., Schorb, S., Rupp, D., Adolph, M., Gorkhover, T., Andersson, I., Hirsemann, H., Potdevin, G., Graafsma, H., Nilsson, B. & Spence, J. C. H. (2011). Nature, 470, 73–77.

Darmanin, C., Strachan, J., Adda, C., Ve, T., Kobe, B. & Abbey, B. (2016). Sci. Rep.6, 25345. PubMed PMC

Egerton, R. F., Li, P. & Malac, M. (2004). Micron, 35, 399–409. PubMed

Fischer, H., Polikarpov, I. & Craievich, A. F. (2004). Protein Sci.13, 2825–2828. PubMed PMC

Fukuta, M., Kanamori, S., Furukawa, T., Nawa, Y., Inami, W., Lin, S., Kawata, Y. & Terakawa, S. (2015). Sci. Rep.5, 16068. PubMed PMC

Fukuta, M., Ono, A., Nawa, Y., Inami, W., Shen, L., Kawata, Y. & Terekawa, S. (2017). J. Biophotonics, 10, 503–510. PubMed

Inami, W., Nakajima, K., Miyakawa, A. & Kawata, Y. (2010). Opt. Express, 18, 12897–12902. PubMed

Johansson, L. C., Stauch, B., Ishchenko, A. & Cherezov, V. (2017). Trends Biochem. Sci.42, 749–762. PubMed PMC

Kaz, D. M., Bischak, C. G., Hetherington, C. L., Howard, H. H., Marti, X., Clarkson, J. D., Adamo, C., Schlom, D. G., Ramesh, R., Aloni, S., Ogletree, D. F. & Ginsberg, N. S. (2013). ACS Nano, 7, 10397–10404. PubMed

Khakurel, K. P., Angelov, B. & Andreasson, J. (2019). Molecules, 24, 3490. PubMed PMC

Kolchiba, M., Inami, W. & Kawata, Y. (2019). Opt. Mater. Expr.9, 1803–1814.

Lunde, C. S., Rouhani, S., Remis, J. P., Ruzin, S. E., Ernst, J. A. & Glaeser, R. M. (2005). J. Appl. Cryst.38, 1031–1034.

Madden, J. T., DeWalt, E. L. & Simpson, G. J. (2011). Acta Cryst. D67, 839–846. PubMed PMC

Maruyama, Y., Ebihara, T., Nishiyama, H., Konyuba, Y., Senda, M., Numaga-Tomita, T., Senda, T., Suga, M. & Sato, C. (2012). Int. J. Mol. Sci.13, 10553–10567. PubMed PMC

Mehrabi, P., Bücker, R., Bourenkov, G., Ginn, H. M., von Stetten, D., Müller-Werkmeister, H. M., Kuo, A., Morizumi, T., Eger, B. T., Ou, W., Oghbaey, S., Sarracini, A., Besaw, J. E., Paré-Labrosse, O., Meier, S., Schikora, H., Tellkamp, F., Marx, A., Sherrell, D. A., Axford, D., Owen, R. L., Ernst, O. P., Pai, E. F., Schulz, E. C. & Miller, R. J. D. (2021). Sci. Adv.7, eabf1380. PubMed PMC

Nannenga, B. L. & Gonen, T. (2019). Nat. Methods, 16, 369–379. PubMed PMC

Nawa, Y., Inami, W., Chiba, A., Ono, A., Miyakawa, A., Kawata, Y., Lin, S. & Terakawa, S. (2012). Opt. Express, 20, 5629. PubMed

Nawa, Y., Inami, W., Lin, S., Kawata, Y. & Terakawa, S. (2015). Opt. Express, 23, 14561. PubMed

Nollert, P. (2003). J. Appl. Cryst.36, 1295–1296. PubMed PMC

Ruoho, M., Valset, K., Finstad, T. & Tittonen, I. (2015). Nanotechnology, 26, 195706. PubMed

Shi, D., Nannenga, B. L., Iadanza, M. G. & Gonen, T. (2013). eLife, 2, e01345. PubMed PMC

Stevenson, H. P., Makhov, A. M., Calero, M., Edwards, A. L., Zeldin, O. B., Mathews, I. I., Lin, G., Barnes, C. O., Santamaria, H., Ross, T. M., Soltis, S. M., Khosla, C., Nagarajan, V., Conway, J. F., Cohen, A. E. & Calero, G. (2014). Proc. Natl Acad. Sci. USA, 111, 8470–8475. PubMed PMC

Wampler, R. D., Kissick, D. J., Dehen, C. J., Gualtieri, E. J., Grey, J. L., Wang, H. F., Thompson, D. H., Cheng, J. X. & Simpson, G. J. (2008). J. Am. Chem. Soc.130, 14076–14077. PubMed PMC