Abstract
Bacteria use complex transporters to secrete functionally relevant proteins to the extracellular medium. The type 2 secretion system (T2SS) translocates folded proteins involved in bacterial nutrient acquisition, virulence and adaptation. The T2SS pseudopilus is a periplasmic filament, assembled by the polymerization of PulG subunits, the major pseudopilin. Pseudopilin proteins have a conserved N-terminal hydrophobic segment followed by a more variable C-terminal periplasmic and globular domain. To better understand the mechanism of assembly and function of the T2SS, we have been studying the structure and dynamics of PulG by NMR, as well as its interaction with other components of the secretion machinery. As a first step on this study, here we reported the chemical shift assignments of PulG C-terminal domain and its secondary structure prediction based on NMR data.
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References
Cavanagh J, Fairbrother WJ, Palmer AG, Skelton NJ (1996) Protein NMR spectroscopy, principles and practice. Academic Press, Inc., San Diego
Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6(3):277–293
Francetic O, Buddelmeijer N, Lewenza S, Kumamoto CA, Pugsley AP (2007) Signal recognition particle-dependent inner membrane targeting of the PulG pseudopilin component of a type II secretion system. J Bacteriol 189(5):1783–1793
Kohler R, Schafer K, Muller S, Vignon G, Diederichs K, Philippsen A, Ringler P, Pugsley AP, Engel A, Welte W (2004) Structure and assembly of the pseudopilin PulG. Mol Microbiol 54(3):647–664
Korotkov K, Sandkvist M, Hol WG (2012) The type II secretion system: biogenesis, molecular architecture and mechanism. Nat Rev Microbiol 10(5):336–351
Nivaskumar M, Francetic O (2014) Type II secretion system: a magic beanstalk or a protein escalator. Biochim Biophys Acta 1843(8):1568–1577
Shen Y, Bax A (2013) Protein backbone and sidechain torsion angles predicted from NMR chemical shifts using artificial neural networks. J Biomol NMR 56(3):227–241
Vranken WF, Boucher W, Stevens TJ, Fogh RH, Pajon A, Llinas M, Ulrich EL, Markley JL, Ionides J, Laue ED (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins 59(4):687–696
Wishart DS, Bigam CG, Yao J, Abildgraad F, Dyson HJ, Oldfield E, Markley JL, Sykes BD (1995) 1H, 13C and 15N chemical shift referencing in biomolecular NMR. J Biomol NMR 6(2):135–140
Yamazaki T, Forman-Kay JD, Kay LE (1993) Two-dimensional NMR experiments for correlating carbon-13 beta and proton delta/epsilon chemical shifts of aromatic residues in 13C-labeled proteins via scalar couplings. J Am Chem Soc 115(23):11054–11055
Acknowledgements
This work was funded by the Institut Pasteur, the Centre National de la Recherche Scientifique (CNRS) and a FiberSpace Grant (No. ANR-14-CE09-0004) from the Agence Nationale de la Recherche (ANR). ALC was funded by the ANR grant. We thank Catherine Simenel for expert help in setting up the NMR experiments. Financial support from the TGIR-RMN-THC Fr3050 CNRS is also gratefully acknowledged.
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López-Castilla, A., Vitorge, B., Khoury, L. et al. 1H, 15N and 13C resonance assignments and secondary structure of PulG, the major pseudopilin from Klebsiella oxytoca type 2 secretion system. Biomol NMR Assign 11, 155–158 (2017). https://doi.org/10.1007/s12104-017-9738-7
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DOI: https://doi.org/10.1007/s12104-017-9738-7