pHluorin-assisted expression, purification, crystallization and X-ray diffraction data analysis of the C-terminal domain of the HsdR subunit of the Escherichia coli type I restriction-modification system EcoR124I

. 2016 Sep ; 72 (Pt 9) : 672-6. [epub] 20160809

Jazyk angličtina Země Spojené státy americké Médium print-electronic

Typ dokumentu časopisecké články

Perzistentní odkaz   https://www.medvik.cz/link/pmid27599856

The HsdR subunit of the type I restriction-modification system EcoR124I is responsible for the translocation as well as the restriction activity of the whole complex consisting of the HsdR, HsdM and HsdS subunits, and while crystal structures are available for the wild type and several mutants, the C-terminal domain comprising approximately 150 residues was not resolved in any of these structures. Here, three fusion constructs with the GFP variant pHluorin developed to overexpress, purify and crystallize the C-terminal domain of HsdR are reported. The shortest of the three encompassed HsdR residues 887-1038 and yielded crystals that belonged to the orthorhombic space group C2221, with unit-cell parameters a = 83.42, b = 176.58, c = 126.03 Å, α = β = γ = 90.00° and two molecules in the asymmetric unit (VM = 2.55 Å(3) Da(-1), solvent content 50.47%). X-ray diffraction data were collected to a resolution of 2.45 Å.

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Csefalvay, E., Lapkouski, M., Guzanova, A., Csefalvay, L., Baikova, T., Shevelev, I., Bialevich, V., Shamayeva, K., Janscak, P., Kuta Smatanova, I., Panjikar, S., Carey, J., Weiserova, M. & Ettrich, R. (2015). PLoS One, 10, e0128700. PubMed PMC

Davies, G. P., Martin, I., Sturrock, S. S., Cronshaw, A., Murray, N. E. & Dryden, D. T. F. (1999). J. Mol. Biol. 290, 565–579. PubMed

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

Dryden, D. T. F., Murray, N. E. & Rao, D. N. (2001). Nucleic Acids Res. 29, 3728–3741. PubMed PMC

Horiuchi, K. & Zinder, N. D. (1972). Proc. Natl Acad. Sci. USA, 69, 3220–3224. PubMed PMC

Janscak, P., Abadjieva, A. & Firman, K. (1996). J. Mol. Biol. 257, 977–991. PubMed

Kabsch, W. (2010). Acta Cryst. D66, 133–144. PubMed PMC

Kobe, B., Ve, T. & Williams, S. J. (2015). Acta Cryst. F71, 861–869. PubMed PMC

Lapkouski, M., Panjikar, S., Janscak, P., Kuta Smatanova, I., Carey, J., Ettrich, R. & Csefalvay, E. (2009). Nature Struct. Mol. Biol. 16, 94–95. PubMed

Loenen, W. A. M., Dryden, D. T. F., Raleigh, E. A. & Wilson, G. G. (2014). Nucleic Acids Res. 42, 20–44. PubMed PMC

Matthews, B. W. (1968). J. Mol. Biol. 33, 491–497. PubMed

Miesenböck, G., De Angelis, D. A. & Rothman, J. E. (1998). Nature (London), 394, 192–195. PubMed

Obarska-Kosinska, A., Taylor, J. E., Callow, P., Orlowski, J., Bujnicki, J. M. & Kneale, G. G. (2008). J. Mol. Biol. 376, 438–452. PubMed PMC

Qi, D. & Scholthof, K. B. (2008). J. Virol. Methods, 149, 85–90. PubMed

Seidel, R., Bloom, J. G., Dekker, C. & Szczelkun, M. D. (2008). EMBO J. 27, 1388–1398. PubMed PMC

Smyth, D. R., Mrozkiewicz, M. K., McGrath, W. J., Listwan, P. & Kobe, B. (2003). Protein Sci. 12, 1313–1322. PubMed PMC

Thor, J. J. van, Georgiev, G. Y., Towrie, M. & Sage, J. T. (2005). J. Biol. Chem. 280, 33652–33659. PubMed

Uyen, N. T., Park, S.-Y., Choi, J.-W., Lee, H.-J., Nishi, K. & Kim, J.-S. (2009). Nucleic Acids Res. 37, 6960–6969. PubMed PMC

Weiss, M. S. & Hilgenfeld, R. (1997). J. Appl. Cryst. 30, 203–205.

Weiss, M. S., Metzner, H. J. & Hilgenfeld, R. (1998). FEBS Lett. 423, 291–296. PubMed

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

Yanisch-Perron, C., Vieira, J. & Messing, J. (1985). Gene, 33, 103–119. PubMed

Youell, J. & Firman, K. (2008). Microbiol. Mol. Biol. Rev. 72, 365–377. PubMed PMC

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