Backbone Assignment of Phosphorylated Cytoplasmic Domain B of Mannitol Transporter IIMtl in Thermoanaerobacter Tengcongensis

  • Lee, Ko On (Department of Agricultural Biotechnology, Seoul National University) ;
  • Suh, Jeong-Yong (Department of Agricultural Biotechnology, Seoul National University)
  • Received : 2017.02.01
  • Accepted : 2017.03.10
  • Published : 2017.03.20


The cytoplasmic domains A and B of the mannitol transporter enzyme $II^{Mtl}$ are covalently linked in Escherichia coli, but separately expressed in Thermoanaerobacter Tengcongensis. The phosphorylation of domain B ($TtIIB^{Mtl}$) substantially increases the binding affinity to the domain A ($TtIIA^{Mtl}$) in T. Tengcongensis. To understand the structural basis of the enhanced domain-domain interaction by protein phosphorylation, we obtained NMR backbone assignments of the phospho-$TtIIB^{Mtl}$ using a standard suite of triple resonance experiments. Our results will be useful to monitor chemical shift changes at the active site of phosphorylation and the binding interfaces.



  1. P. W. Postma, J. W. Lengeler, and G. R. Jacobson, Microbiol. Rev. 57, 543 (1993)
  2. C. Siebold, K. Flukiger, R. Beutler, and B. Erni, FEBS Lett. 504, 104 (2001)
  3. J. Deutscher, C. Francke, and P. W. Postma, Microbiol. Mol. Biol. Rev. 70, 939 (2006)
  4. Q. Bao,Y. Tian, W. Li, Z. Xu, Z. Xuan, S. Hu, W. Dong, J. Yang, Y. Chen, Y. Xue, Y. Xu, X. Lai, L. Huang, X. Dong, Y. Ma, L. Ling, H. Tan, R. Chen, J. Wang, J. Yu, and H. Yang, Genome Res. 12, 689 (2002)
  5. P.M. Legler, M.Cai, A. Peterkofsky, and G. M. Clore, J. Biol. Chem. 279, 39115 (2004)
  6. Suh, C. Tang, M. Cai, and G. M. Clore, J. Mol. Biol. 353, 1129 (2005)
  7. J. Y. Suh, M. Cai, and G. M. Clore, J. Biol. Chem. 283, 18980 (2008)
  8. J. Y. Suh, J. Iwahara, and G. M. Clore, Proc. Natl. Acad. Sci. U.S.A. 104, 3153 (2007)
  9. V. P. R. Chichili, V. Kumar, and J. Sivaraman, Protein Sci. 22, 153 (2013)
  10. K. O. Lee, E. H. Kim, G. Kim, J. Y. Jung, S. Katayama, S. Nakamura, and J. Y. Suh, Protein Sci. 25, 1803 (2016)
  11. F. Delaglio, S. Grzesiek, G. W. Vuister, G. Zhu , J. Pfeifer, and A. Bax, J. Biomol. NMR 6, 277 (1995)
  12. D. S. Garrett, R. Powers, A. M. Gronenborn, and G. M. Clore, J. Magn. Reson. 95, 214 (1991)
  13. B. A. Johnson, and R. A. Blevins, J. Biomol. NMR 4, 603 (1994)
  14. M. D. Seo, S. J. Park, S. H. Seok, J. H. Kim, M. J. Cha, and B. J. Lee, J. Kor. Magn. Reson. Soc. 16, 91 (2012)
  15. K. O. Lee, and J. Y. Suh, J. Kor. Magn. Reson. Soc. 19, 42 (2015)
  16. Y. Shen, F. Delaglio, G. Cornilescu, and A. Bax , J. Biomol. NMR 44, 213 (2009)