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Stability and Structure of S128A Mutant cAMP Receptor Protein

  • Received : 2011.07.27
  • Accepted : 2011.08.31
  • Published : 2011.09.30

Abstract

Cyclic AMP receptor protein(CRP) is involved in the activation of many genes corresponding to catabolite enzymes in Escherichia coli. In this study, mutant CRP(S128A) was used to elucidate the effect of Ser 128 on the cAMP-induced structural change. Based on the protease digestion and thermal analysis, serine 128 in CRP affects the cAMP binding capability and then structural change of CRP protein. In addition, CD spectra in near UV region revealed that S128A CRP retained the sensitive conformation to thermal effect relative to that of wild-type CRP, in spite of identical Tm values in the absence of cAMP.

Keywords

References

  1. deCrombrugghe, S. Busby, and Buc, "CAMP receptor protein: role in transcription activation", Science, Vol. 224, p. 831, 1984. https://doi.org/10.1126/science.6372090
  2. W. S. Reznikoff, "Catabolite gene activator protein activation of lac transcription", J. Bacteriol., Vol. 174, p. 655, 1992. https://doi.org/10.1128/jb.174.3.655-658.1992
  3. S. Garges and S. Adhya, "Sites of allosteric shift in the structure of the cyclic AMP receptor protein", Cell, Vol. 41, p. 745, 1985. https://doi.org/10.1016/S0092-8674(85)80055-6
  4. J. G. Harman, "Allosteric regulation of the cAMP receptor protein", Biochimica et Biophysica Acta, Vol. 1547, p. 1, 2001. https://doi.org/10.1016/S0167-4838(01)00187-X
  5. J. Malecky, A. Polit, and Z. Wasylewski, "Kinetic studies of cAMP-induced allosteric changes in cyclic AMP receptor protein from Escherichia coli", J. Biol. Chem., Vol. 275, p. 8480, 2000. https://doi.org/10.1074/jbc.275.12.8480
  6. J. M. Passner and T. A. Steitz, "The structure of a CAP-DNA complex having two cAMP molecules bound to each monomer", Proc. Natl. Acad. Sci. USA, Vol. 94, p. 2843, 1997. https://doi.org/10.1073/pnas.94.7.2843
  7. J. Mukhopadhyay, R. Sur, and P. Parrack, "Functional roles of the cyclic AMP-dependent forms of cyclic AMP receptor protein from Escherichia coli", FEBS Lett., Vol. 453, p. 215, 1999. https://doi.org/10.1016/S0014-5793(99)00719-X
  8. E. J. Lee, J. Glasgow, S. F. Leu, A. O. Belduz, and J. G. Harman, "Mutagenesis of the cyclic AMP receptor protein of Escherichia coli", Nucleic Acids Res., Vol. 22, p. 2894, 1994. https://doi.org/10.1093/nar/22.15.2894
  9. J. B. Gang, "Measurement of DNA helical change for the binding of cyclic AMP receptor protein to lac DNA", Biochemical and Biophysical Research Communications, Vol. 322, p. 993, 2004. https://doi.org/10.1016/j.bbrc.2004.08.011
  10. J. G. Harman, A. Peterkofsky, and K. McKenney, "Arginine substituted for leucine at position 195 produces a cyclic AMP-independent form of Escherichia coli", J. Biol. Chem., Vol. 263, p. 8072, 1988.
  11. J. B. Gang, and J. G. Harman, "The characteristic binding of mutant cyclic AMP receptor protein to lactose promoter in Escherichia coli", Mol Cells, Vol. 4, p. 167, 1994.
  12. J. B. Gang, H. J. Chung, G. G. Park, and Y. S. Park, "Stability and structural change of cAMP receptor protein at low and high cAMP concentrations", J. Microbiol. Biotechnol., Vol. 15, p. 1392, 2005.
  13. X. Cheng, L. Kovac, and J. C. Lee, "Probing the mechanism of CRP activation by site-directed mutagenesis", Biochemistry, Vol. 34, p. 10816, 1995. https://doi.org/10.1021/bi00034a014
  14. M. M. Santoro and D. W. Bolen, "Unfolding free energy changes determined by the linear extrapolation method", Biochemistry, 27, p. 8063, 1988. https://doi.org/10.1021/bi00421a014
  15. A. Polit, P. Bonarek, B. Kepys, S. Kedracka-Krok, A. Gorecki, and Z. Wasylewski, "Kinetic studies of cAMP-induced allosteric changes in mutants T127I, S128A, and T127I/S128A of the cAMP receptor protein from Escherichia coli", J. Biol. Chem., Vol. 278, p. 43020, 2003. https://doi.org/10.1074/jbc.M306398200
  16. A. Polit, U. Blaszczyk, and Z. Wasylewski, "Steadystate and time-resolved fluorescence studies of conformational change induced by cyclic AMP and DNA binding to cyclic AMP receptor protein from Escherichia coli", Eur. J. Biochem., 270, p. 1413, 2003. https://doi.org/10.1046/j.1432-1033.2003.03497.x
  17. J. H. Joo, M.-J. Goo, and J. Gang, "Study on the structure of cAMP receptor protein (CRP) by temperature change", Kor. J. Life Science, Vol. 10, p. 279, 2000.
  18. J. B. Gang and H. Y. Cho, "Application of temperature gradient gel electrophoresis to cAMP receptor protein", J. Life. Science, Vol. 14, p. 309, 2004. https://doi.org/10.5352/JLS.2004.14.2.309