Biomolecular NMR Assignments

, Volume 3, Issue 1, pp 61–65

1H, 15N, 13C resonance assignments of the reduced and active form of human Protein Tyrosine Phosphatase, PRL-1

Article

Abstract

Phosphatase of regenerating liver-1 (PRL-1) is a novel target for potentially treating cancer metastases. Although its specific biochemical role in these processes has yet to be delineated, considerable evidence suggests the phosphatase activity of PRL-1 is required for promoting cancer and metastasis. PRL-1 belongs to the protein tyrosine phosphatase (PTPase) family and functions using the CX5R consensus active site motif. Like other PTPases, PRL-1 is inhibited by oxidation at its active site Cys, however, disulfide bond formation occurs unusually readily in wild-type PRL-1. Chemical shift assignments are available for oxidized wild type, but numerous, substantial changes are observed in the spectra upon reduction. Because the reduced form is active, we sought to identify a stable mutant that would resist oxidation and be useful for facilitating drug screening and development using NMR-based assays. We present here NMR assignments for a full-length, reduced and active form of PRL-1, PRL-1-C170S-C171S, that is well suited for this purpose.

Keywords

PRL-1 PTPase Resonance assignments Oxidation Reduction Redox Drug screening 

References

  1. Delaglio F, Grzesiek S et al (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293. doi:10.1007/BF00197809 CrossRefGoogle Scholar
  2. Eghbalnia HR, Bahrami A et al (2005) Probabilistic identification of spin systems and their assignments including coil-helix interference as output (PISTACHIO). J Biomol NMR 32:219–233. doi:10.1007/s10858-005-7944-6 CrossRefGoogle Scholar
  3. Goddard TD, Kneller DG (2004) SPARKY 3. University of California, San FranciscoGoogle Scholar
  4. Jeong DG, Kim SJ et al (2005) Trimeric structure of the PRL-1 phosphatase reveals an active enzyme conformation and regulation mechanisms. J Mol Biol 345:401–413. doi:10.1016/j.jmb.2004.10.061 CrossRefGoogle Scholar
  5. Kozlov G, Cheng J et al (2002) Letter to the Editor: 1H, 13C and 15N resonance assignments of the human phosphatase PRL-3. J Biomol NMR 24:169–170. doi:10.1023/A:1020937316065 CrossRefGoogle Scholar
  6. Kozlov G, Cheng J et al (2004) Structural insights into molecular function of the metastasis-associated phosphatase PRL-3. J Biol Chem 279:11882–11889. doi:10.1074/jbc.M312905200 CrossRefGoogle Scholar
  7. Laurence JS, Hallenga K et al (2004) Letter to the Editor: 1H, 15N, 13C resonance assignments of the human protein tyrosine phosphatase PRL-1. J Biomol NMR 29:417–418. doi:10.1023/B:JNMR.0000032506 CrossRefGoogle Scholar
  8. Sharma D, Rajarathnam K (2000) 13C NMR chemical shifts can predict disulfide bond formation. J Biomol NMR 18:165–171. doi:10.1023/A:1008398416292 CrossRefGoogle Scholar
  9. Stephens BJ, Han H et al (2005) PRL phosphatases as potential molecular targets in cancer. Mol Cancer Ther 4:1653–1661. doi:10.1158/1535-7163.MCT-05-0248 CrossRefGoogle Scholar
  10. Sun J-P, Wang W-Q et al (2005) Structure and biochemical properties of PRL-1, a phosphatase implicated in cell growth, differentiation, and tumor invasion. Biochemistry 44:12009–12021. doi:10.1021/bi0509191 CrossRefGoogle Scholar
  11. Wishart DS, Bigam CG et al (1995) 1H, 13C, and 15N chemical shift referencing in biomolecular NMR. J Biomol NMR 6:135–140. doi:10.1007/BF00211777 CrossRefGoogle Scholar
  12. Zhou H, Gallina M et al (2003) Letter to the Editor: 1H, 13C, and 15N resonance assignments and secondary structure of the human protein tyrosine phosphatase PRL-2. J Biomol NMR 27:397–398. doi:10.1023/A:1025875618084 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  1. 1.Department of Pharmaceutical ChemistryThe University of KansasLawrenceUSA

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