[en] The capillary counterdiffusion method is a very efficient crystallization technique for obtaining high-quality protein crystals. This technique requires a convection-free environment, which can be achieved using either gelled solutions, very thin capillaries, or microgravity conditions. To study the influence of a convection-free environment on protein crystal quality and to evaluate two different experimental implementations to achieve it, we have made a comparative analysis of crystals grown by capillary counterdiffusion in agarose, a convective-free environment on Earth, and crystals grown in microgravity at the International Space Station. Thermotoga maritima triose phosphate isomerase (TIM) was chosen as a model for this study. The statistical analysis reveals a significant improvement for the crystals grown in microgravity in terms of their R-merge, B-value, and mosaicity, but the statistical evidence is insufficient to show a similar benefit for the resolution and mean intensity parameters. These results are quite surprising because it is known that, unlike gels, the noisy microgravity scenario offered by the ISS cannot sustain a convection-free environment on the time scale of days required for protein crystallization experiments.
Beaucamp, N., Hofmann, A., Kellerer, B., Jaenicke, R., Dissection of the gene of the bifunctional PGK-TIM fusion protein from the hyperthermophilic bacterium Thermotoga maritima: Design and characterization of the separate triosephosphate isomerase (1997) Protein Sci, 6, pp. 2159-2165
Boggon, T.J., Chayen, N.E., Snell, E.H., Dong, J., Lautenschlager, P., Potthast, L., Siddons, D.P., Helliwell, J.R., Protein crystal movements and fluid flows during microgravity growth (1998) Philos. Trans. R. Soc. London, Ser. A, 356, pp. 1045-1061
Chayen, N.E., Saridakis, E., Protein crystallization for genomics: Towards high-throughput optimization techniques (2002) Acta Crystallogr., Sect. D, 58, pp. 921-927
Chayen, N.E., Turning protein crystallisation from an art into a science (2004) Curr. Opin. Struct. Biol, 14, pp. 577-583
Garcia-Ruiz, J.M., Moreno, A., Viedma, C., Coll, M., Crystal quality of lysozyme single crystals grown by the gel acupuncture method (1993) Mater. Res. Bull, 28, pp. 541-546
Garcia-Ruiz, J.M., Moreno, A., Investigations on protein crystal growth by the gel acupuncture method (1994) Acta Crystallogr., Sect. D, 50, pp. 484-490
García-Ruiz, J.M., Otálora, F., Crystal growth studies in microgravity with the APCF. II. Image analysis studies (1997) J. Cryst. Growth, 182, pp. 155-167
Garcia-Ruiz, J.M., Gavira, J.A., Otalora, F., Guasch, A., Coll, M., Reinforced protein crystals (1998) Mater. Res. Bull, 33, pp. 1593-1598
Garcia-Ruiz, J.M., Otalora, F., Novella, M.L., Gavira, J.A., Sauter, C., Vidal, O., A supersaturation wave of protein crystallization (2001) J. Cryst. Growth, 232, pp. 149-155
García-Ruiz, J.M., Drenth, J., Ries-Kautt, M., Tardieu, A., (2001) A World without Gravity: Research in Space for Health and Industrial Processes, 1251, pp. 150-171. , Seibert, G, Ed, European Space Agency: Paris
Garcia-Ruiz, J.M., Gonzalez-Ramirez, L.A., Gavira, J.A., Otalora, F., Granada Crystallisation Box: A new device for protein crystallisation by counter-diffusion techniques (2002) Acta Crystallogr., Sect. D, 58, pp. 1638-1642
Gavira, J.A., Garcia-Ruiz, J.M., Agarose as crystallisation media for proteins II: Trapping of gel fibres into the crystals (2002) Acta Crystallogr., Sect. D, 58, pp. 1653-1656
Judge, R.A., Snell, E.H., van der Woerd, M.J., Extracting trends from two decades of microgravity macromolecular crystallization history (2005) Acta Crystallogr., Sect. D, 61, pp. 763-771
Kabsch, W.J., Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants (1993) J. Appl. Crystallogr, 26, pp. 795-800
Kundrot, C.E., Judge, R.A., Pusey, M.L., Snell, E.H., Microgravity and macromolecular crystallography (2001) Cryst. Growth Des, 1, pp. 87-99
Maes, D., Zeelen, J.P., Thanki, N., Beaucamp, N., Alvarez, M., Thi, M.H., Backmann, J., Wierenga, R.K., The crystal structure of triosephosphate isomerase (TIM) from Thermotoga maritima: A comparative thermostability structural analysis of ten different TIM structures (1999) Proteins, 37, pp. 441-453
Matsumoto, S., Yoda, S.J., Numerical study of diffusion coefficient measurements with sinusoidal varying accelerations (1999) J. Appl. Phys, 85, pp. 8131-8136
Mcpherson, A., Macromolecular crystal growth in microgravity (1999) Crystallogr. Rev, 6, pp. 157-305
Otálora, F., Garcia-Ruiz, J.M., Crystal growth studies in microgravity with the APCF I. Computer simulation of transport dynamics (1997) J. Cryst. Growth, 182, pp. 141-154
Otwinowski, Z., Minor, W., Processing of X-ray diffraction data collected in oscillation mode (1997) Methods Enzymol, 276, pp. 307-326
Pusey, M., Witherow, W.K., Nauman, R., Preliminary investigations into solutal flow about growing tetragonal lysozyme crystals (1988) J. Cryst. Growth, 90, pp. 105-111
Snell, E.H., Boggon, T.J., Helliwell, J.R., Moskowitz, M.E., Nadarajah, A., CCD video observation of microgravity crystallization of lysozyme and correlation with accelerometer data (1997) Acta Crystallogr., Sect. D, 53, pp. 747-755
Snell, E.H., Helliwell, J.R., Macromolecular crystallization in microgravity (2005) Rep. Prog. Phys, 68, pp. 799-853
Vergara, A., Lorber, B., Zagari, A., Giege, R., Physical aspects of protein crystal growth investigated with the Advanced Protein Crystallization Facility in reduced-gravity environments (2003) Acta Crystallogr, Sect. D, 59, pp. 2-15
Zegers, I., Carotenuto, L., Evrard, C., Garcia-Ruiz, J.M., De Gieter, P., Gonzalez-Ramirez, L.A., Istasse, E., Dubois, F., Counter-diffusion protein crystallisation in microgravity and its observation with PromlSS (Protein Microscope for the International Space Station) (2006) Microgravity Sci. Technol, 23, pp. 165-169
Zegers, I., Van De Weerdt, C., Evrard, C., Vanhee, C., Sleutel, M., Willaert, R., Declercq, J.P., Garcia-Ruiz, J.M., (2007), unpublished work
