Artículos de revistas
Crystallization And Preliminary X-ray Diffraction Analysis Of Q4dv70 From Trypanosoma Cruzi, A Hypothetical Protein With A Putative Thioredoxin Domain
Registro en:
Acta Crystallographica Section F: Structural Biology And Crystallization Communications. , v. 65, n. 6, p. 641 - 644, 2009.
17443091
10.1107/S1744309109017734
2-s2.0-67649227258
Autor
Dos Santos C.R.
Fessel M.R.
Vieira L.D.C.
Krieger M.A.
Goldenberg S.
Guimaraes B.G.
Zanchin N.I.T.
Barbosa J.A.R.G.
Institución
Resumen
Q4DV70 is annotated in the Trypanosoma cruzi CL Brener genome as a hypothetical protein with a predicted thioredoxin-like fold, although the catalytic cysteine residues that are conserved in typical oxidoreductases are replaced by serine residues. Gene-expression analysis indicates that this protein is differentially expressed during the T. cruzi life cycle, suggesting that it plays an important role during T. cruzi development. The gene coding for Q4DV70 was cloned and the protein was overexpressed in Escherichia coli with an N - terminal His tag. Purification of Q4DV70 was carried out by affinity and size-exclusion chromatography and the His tag was removed by TEV protease digestion. Crystals of Q4DV70 were grown using the sitting-drop vapour-diffusion method. A diffraction data set was collected to 1.50 Å resolution from a single crystal grown in 25% PEG 1500, 200 mM sodium thiocyanate pH 6.9, 10 mM phenol and 10% ethylene glycol. The crystal belonged to space group P212121, with unit-cell parameters a = 35.04, b = 50.32, c = 61.18 Å. The Q4DV70 structure was solved by molecular replacement using protein disulfide isomerase from yeast (PDB code 2b5e) as a search model. Initial refinement of the model indicated that the solution was correct. These data are being used for refinement of the model of Q4DV70. © 2009 International Union of Crystallography. All rights reserved. 65 6 641 644 Avila, A.R., Dallagiovanna, B., Yamada-Ogatta, S.F., Monteiro-Góes, V., Fragoso, S.P., Krieger, M.A., Goldenberg, S., (2003) Genet. Mol. Res, 2, pp. 159-168 Brünger, A.T., Adams, P.D., Clore, G.M., DeLano, W.L., Gros, P., Grosse-Kunstleve, R.W., Jiang, J.-S., Warren, G.L., (1998) Acta Cryst, D54, pp. 905-921 Cai, H., Wang, C.C., Tsou, C.L., (1994) J. Biol. Chem, 269, pp. 24550-24552 Carneiro, F.R.G., Silva, T.L.C., Alves, A.C., Haline-Vaz, T., Gozzo, F.C., Zanchin, N.I., (2006) Biochem. Biophys. Res. Commun, 3 (43), pp. 260-268 Chagas, C., (1909) Mem. Inst. Oswaldo Cruz, 1, pp. 159-218 Chen, J., Song, J.L., Zhang, S., Wang, Y., Cui, D.F., Wang, C.C., (1999) J. Biol. Chem, 274, pp. 19601-19605 Clayton, C., Shapira, M., (2007) Mol. Biochem. Parasitol, 156, pp. 93-101 De Souza, W., (1984) Int. Rev. Cytol, 86, pp. 197-283 Emanuelsson, O., Brunak, S., von Heijne, G., Nielsen, H., (2007) Nature Protoc, 2, pp. 953-971 Emsley, P., Cowtan, K., (2004) Acta Cryst, D60, pp. 2126-2132 Epp, O., Ladenstein, R., Wendel, A., (1983) Eur. J. Biochem, 133, pp. 51-69 Gilliland, G.L., Tung, M., Blakeslee, D.M., Ladner, J.E., (1994) Acta Cryst, D50, pp. 408-413 Haile, S., Papadopoulou, B., (2007) Curr. Opin. Microbiol, 10, pp. 569-577 Hol, W.G.J., Sarfaty, S.H., Stewart, L.J., Kim, H., (2001), US Patent 6267935Holmgren, A., Söderberg, B.-O., Eklund, H., Brändén, C.-I., (1975) Proc. Natl Acad. Sci. USA, 72, pp. 2305-2309 Jancarik, J., Kim, S.-H., (1991) J. Appl. Cryst, 24, pp. 409-411 Kanaujia, S.P., Ranjani, C.V., Jeyakanthan, J., Baba, S., Chen, L., Liu, Z.-J., Wang, B.-C., Yokoyama, S., (2007) Acta Cryst, F63, pp. 27-29 Kemmink, J., Darby, N.J., Dijkstra, K., Nilges, M., Creighton, T.E., (1996) Biochemistry, 35, pp. 7684-7691 Kern, R., Malki, A., Holmgren, A., Richarme, G., (2003) Biochem. J, 371, pp. 965-972 Majeed, S., Ofek, G., Belachew, A., Huang, C.C., Zhou, T., Kwong, P.D., (2003) Structure, 11, pp. 1061-1070 Martin, J.L., (1995) Structure, 3, pp. 245-250 Martin, J.L., Bardwell, J.C.A., Kuriyan, J., (1993) Nature (London), 365, pp. 464-468 Matthews, B.W., (1968) J. Mol. Biol, 33, pp. 491-497 Minning, T.A., Bua, J., Garcia, G.A., McGraw, R.A., Tarleton, R.L., (2003) Mol. Biochem. Parasitol, 131, pp. 55-64 Murshudov, G.N., Vagin, A.A., Dodson, E.J., (1997) Acta Cryst, D53, pp. 240-255 Newman, J., Egan, D., Walter, T.S., Meged, R., Berry, I., Ben Jelloul, M., Sussman, J.L., Perrakis, A., (2005) Acta Cryst, D61, pp. 1426-1431 Otwinowski, Z., Minor, W., (1997) Methods Enzymol, 276, pp. 307-326 Page, R., Grzechnik, S.K., Canaves, J.M., Spraggon, G., Kreusch, A., Kuhn, P., Stevens, R.C., Lesley, S.A., (2003) Acta Cryst. D, 59, pp. 1028-1037 Reinemer, P., Dirr, H.W., Ladenstein, R., Schäffer, J., Gallay, O., Huber, R., (1991) EMBO J, 10, pp. 1997-2005 Sodano, P., Xia, T.H., Bushweller, J.H., Bjö rnberg, O., Holmgren, A., Billeter, M., Wüthrich, K., (1991) J. Mol. Biol, 221, pp. 1311-1324 Stein, N., (2008) J. Appl. Cryst, 41, pp. 641-643 Vagin, A., Teplyakov, A., (1997) J. Appl. Cryst, 30, pp. 1022-1025 Zheng, W.D., Quan, H., Song, J.L., Yang, S.L., Wang, C.C., (1997) Arch. Biochem. Biophys, 337, pp. 326-331