Biomolecular NMR Assignments

, Volume 13, Issue 1, pp 239–243 | Cite as

Backbone and side chain 1H, 15N and 13C assignments of a putative peptidyl prolyl cis–trans isomerase FKBP12 from Mycobacterium tuberculosis

  • Guilherme Caldas Andrade
  • Luis Felipe Correa Silva
  • Danielle Maria Perpétua Oliveira
  • José Ricardo M. Pires
  • Fabio C. L. AlmeidaEmail author
  • Cristiane Dinis AnobomEmail author


FK506 Binding Proteins (FKBPs) are a family of highly conserved and important proteins that possess a peptidyl cis–trans isomerase (PPIases) domain. Human FKBP12 is a prototype of this family and it is involved in many diseases due to its interaction with the immunosuppressive drugs FK506 and rapamycin. They inhibit calcineurin and mTOR complex, respectively, leading to parasite death by inhibiting cell proliferation through cytokinesis blockade being an important target to find new drugs. Tuberculosis is a disease that causes important impacts on public health worldwide. In this context, MtFKBP12 is a putative peptidyl prolyl cis–trans isomerase from Mycobacterium tuberculosis and here we report the NMR chemical shift assignment for 1H, 15N and 13C nuclei in the backbone and side chains of the MtFKBP12. This lays the foundation for further structural studies, backbone dynamics, mapping of interactions and drug screening and development. We have found through the NMR spectrum that the protein is well folded with narrow peaks and almost none overlap in 15N-HSQC. Prediction of secondary structure using Talos-N server showed great similarity with other proteins from this family.


FKBP Mycobacterium tuberculosis NMR Assignment 



This work was funded by FAPERJ Grants 239229 and 204432, awarded to FCLA, CNPq Grant 309564/2017-4, awarded to FCLA. We also thank to INBEB-INCT for funding. The assignment was deposit at the Biomagnetic Resonance Data Bank (BMRB ID 27690). GCA is funded by CAPES scholarship and LFCS is funded by FAPERJ.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


  1. Aido-Machado R, Salmon D, Pires JR (2014) 1H, 15N and 13C assignments of a putative peptidyl prolyl cis-trans isomerase FKBP12 from Trypanosoma brucei. Biomol NMR Assign 8:133–135. CrossRefGoogle Scholar
  2. Brasseur A, Rotureau B, Vermeersch M et al (2013) Trypanosoma brucei FKBP12 differentially controls motility and cytokinesis in procyclic and bloodstream forms. Eukaryot Cell 12:168–181. CrossRefGoogle Scholar
  3. Delaglio F, Grzesiek S, Vuister GW et al (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293. CrossRefGoogle Scholar
  4. Gal M, Edmonds KA, Milbradt AG et al (2012) Speeding up direct 15 N detection: hCaN 2D NMR experiment Maayan. J Biomol NMR 51:497–504. CrossRefGoogle Scholar
  5. Galat A (2003) Peptidylprolyl cis/trans isomerases (Immunophilins): biological diversity-targets-functions. Curr Topics Med Chem 3(12):1215–1347CrossRefGoogle Scholar
  6. Greenstein RJ, Su L, Juste RA, Brown ST (2008) On the action of cyclosporine A, rapamycin and tacrolimus on M. avium including subspecies paratuberculosis. PLoS ONE 3:1–6. CrossRefGoogle Scholar
  7. Greenstein RJ, Su L, Shahidi A et al (2014) Unanticipated Mycobacterium tuberculosis complex culture inhibition by immune modulators, immune suppressants, a growth enhancer, and vitamins A and D: clinical implications. Int J Infect Dis 26:37–43. CrossRefGoogle Scholar
  8. Griesinger C, Sattler M (1999) Heteronuclear multidimensional NMR experiments for the structure determination of proteins in solution employing pulsed field gradients. Prog Nuclear Magn Reson Spectrosc 34:93–158Google Scholar
  9. Grzesiek S, Bax A (1993) Amino acid type determination in the sequential assignment procedure of uniformly 13C/15N-enriched proteins. J Biomol NMR 3:185–204Google Scholar
  10. Ikura M, Kay LE, Bax A (1990) A novel-approach for sequential assignment of H-1, C-13, and N-15 spectra of larger proteins—heteronuclear triple-resonance 3-dimensional Nmr-spectroscopy—application to calmodulin. Biochemistry 29:4659–4667CrossRefGoogle Scholar
  11. Kay LE, Xu GY, Singer AU et al (1993) A gradient-enhanced HCCH-TOCSY experiment for recording side-chain 1H and 13C correlations in H2O samples of proteins. J Magn Reson B 101:333–337. ADSCrossRefGoogle Scholar
  12. Liu J, Farmer JD, Lane WS et al (1991) Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell 66:807–815. CrossRefGoogle Scholar
  13. Logan TM, Olejniczak ET, Xu RX, Fesik SW (1992) Side chain and backbone assignments in isotopically labeled proteins from two heteronuclear triple resonance experiments. FEBS Lett 314:413–418CrossRefGoogle Scholar
  14. Moro A, Ruiz-Cabello F, Fernández-Cano A et al (1995) Secretion by Trypanosoma cruzi of a peptidyl-prolyl cis-trans isomerase involved in cell infection. EMBO J 14:2483–2490CrossRefGoogle Scholar
  15. Mustafi SM, Chen H, Li H et al (2013) Analysing the visible conformational substates of the FK506-binding protein FKBP12. Biochem J 453:371–380. CrossRefGoogle Scholar
  16. Pai M, Behr MA, Dowdy D et al (2016) Tuberculosis. Nat Rev Dis Prim. Google Scholar
  17. Shen Y, Bax A (2013) Protein backbone and sidechain torsion angles predicted from NMR chemical shifts using artificial neural networks. J Biomol NMR 56:227–241. CrossRefGoogle Scholar
  18. Shuker SB, Hajduk PJ, Meadows RP, Fesik SW (1996) Discovering high-affinity ligands for proteins: SAR by NMR. Science 274:1531–1534. ADSCrossRefGoogle Scholar
  19. Tong M, Jiang Y (2015) FK506-binding proteins and their diverse functions. Curr Mol Pharmacol. Google Scholar
  20. Vranken WF, Boucher W, Stevens TJ et al (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins Struct Funct Genet 59:687–696. CrossRefGoogle Scholar
  21. WHO (2017) Global Tuberculosis Report 2017Google Scholar
  22. Wittekind M, Mueller L (1993) HNCACB, a high-sensitivity 3D NMR experiment to correlate amide-proton and nitrogen resonances with the alpha- and beta-carbon resonances in proteins. J Magn Reson B 101:201–205. CrossRefGoogle Scholar
  23. Zumla A, Chakaya J, Centis R et al (2015) Tuberculosis treatment and management-an update on treatment regimens, trials, new drugs, and adjunct therapies. Lancet Respir Med 3:220–234. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Institute of ChemistryFederal University of Rio de JaneiroRio de JaneiroBrazil
  2. 2.Institute of Medical Biochemistry (IBqM), National Center of Nuclear Magnetic Resonance Jiri JonasFederal University of Rio de JaneiroRio de JaneiroBrazil
  3. 3.National Center of Nuclear Magnetic Resonance (CNRMN), Center for Structural Biology and Bioimaging (CENABIO)Federal University of Rio de JaneiroRio de JaneiroBrazil

Personalised recommendations