Preparation of the recombinant НU-proteins from S. melliferum and M. gallisepticum and of their complexes with DS-DNA for structural NMR experiments
Tóm tắt
We created a high-performance system for expression of 13C, 15N-double-labeled recombinant НU-proteins from mycoplasms S. melliferum (HUSpm) and M. gallisepticum (HUMgal), and a purification protocol that yields protein samples suitable for structure studies by high-resolution NMR. Using these proteins we optimized the conditions of NMR experiments and obtained two-dimensional heteronuclear 1H/15N-HSQC and 1H/13С-HSQC-CT NMR spectra of free НU-proteins (HUSpm and HUMGal), and 1H/15N-HSQC spectrum of a complex between HUSpm and a double-stranded DNA (ds-DNA). The techniques we created are applicable in the studies of НU-proteins from other organisms. They allow analyzing structure and dynamics of free proteins and of their conformational changes induced by binding of various ds-DNAs or of DNA binding inhibitors.
Tài liệu tham khảo
Drlica, K. and Rouviere-Yaniv, J., Microbiol. Rev., 1987, vol. 51, p. 301.
Dame, R.T., Mol. Microbiol., 2005, vol. 56, p. 858.
Rouviere-Yaniv, J. and Gros, F., Proc. Natl. Acad. Sci. U. S. A., 1975, vol. 72, p. 3428.
Bensaid, A., Almeida, A., Drlica, K., and Rouviere- Yaniv, J., J. Mol. Biol., 1996, vol. 256, p. 292.
Dorman, C.J., Nat. Rev. Microbiol., 2013, vol. 11, p. 349.
Kamashev, D. and Rouviere-Yaniv, J., EMBO J., 2000, vol. 19, p. 6527.
Oberto, J., Nabti, S., Jooste, V., Mignot, H., and Rouviere-Yaniv, J., PLoS One, 2009, p. 4.
Grove, A., Curr. Issues Mol. Biol., 2010, vol. 13, p. 1.
Huisman, O., Faelen, M., Girard, D., Jaffé, A., Toussaint, A., and Rouvière-Yaniv, J., J Bacteriol., 1989, vol. 171, p. 3704.
Dri, A.M., Rouviere-Yaniv, J., and Moreau, P.L., J. Bacteriol., 1991, vol. 173, p. 2852.
Fernandez, S., Rojo, F., Alonso, J.C., Mol. Microbiol., 1997, vol. 23, p. 1169.
Bhowmick, T., Ghosh, S., Dixit, K., Ganesan, V., Ramagopal, U.A., Dey, D., Sarma, S.P., Ramakumar, S., and Nagaraja, V., Nat. Commun., 2014, vol. 5, p. 4124.
Zhang, R. and Lin, Y., Nucleic Acids Res., 2009, vol. 37, p. 455.
Dillon, S.C. and Dorman, C.J., Nat. Rev. Microbiol., 2010, vol. 8, p. 185.
Glass, J.I., Assad-Garcia, N., Alperovich, N., Yooseph, S., Lewis, M.R., Maruf, M., Hutchison, C.A., Smith, H.O., and Venter, J.C., Proc. Natl. Acad. Sci. U. S. A., 2006, vol. 103, p. 425.
Anderson, A., Chem. Biol., 2003, vol. 10, p. 787.
Boyko, K., Gorbacheva, M., Rakitina, T., Korzhenevskiy, D., Vanyushkina, A., Kamashev, D., Lipkin, A., and Popov, V., Acta Crystallogr., Sect. F: Struct. Biol. Commun., 2015, vol. 71, p. 24.
Nikolaeva, A.Yu., Timofeev, V.I., Boiko, K.M., Korzhenevskii, D.A., Rakitina, T.V., Dorovatovskii, P.V., and Lipkin, A.V., Cristallogr. Rep., 2015, vol. 60, no. 6, p. 880.
Cavanagh, J., Fairbrother, W.J., Palmer, A.G., and Skelton, N.J., Protein NMR Spectroscopy: Principles and Practice, San Diego: Academic, 2006.
Studier, F.W., Protein Expression Purif., 2005, vol. 41, p. 207.
Laemmli, U.K., Nature, 1970, vol. 227, no. 5259, p. 680.
Balandina, A., Kamashev, D., and Rouviere-Yaniv, J., J. Biol. Chem., 2002, vol. 277, p. 27622.
Garner, M.M. and Revzin, A.A., Nucleic Acids Res., 1981, vol. 9, p. 3047.
Kamashev, D.E., Esipova, N.G., Ebralidse, K.K., and Mirzabekov, A.D., FEBS Lett., 1995, vol. 375, p. 27.