How thiamine diphosphate is activated in enzymes

Science

Volumn 275, Issue 5296, 1997, Pages 67-70

  Kern, Dorothee ^a   Kern, Gunther ^a   Neef, Holger ^a   Tittmann, Kai ^a   Killenberg Jabs, Margrit ^a   Wikner, Christer ^a   Schneider, Gunter ^a   Hübner, Gerhard ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

COCARBOXYLASE; GLUTAMIC ACID; PYRUVATE DECARBOXYLASE; THIAMINE; TRANSKETOLASE;

ARTICLE; CATALYSIS; CHEMICAL REACTION KINETICS; COENZYME; ENZYME ACTIVATION; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; PRIORITY JOURNAL; PROTON TRANSPORT; THERMODYNAMICS;

ALLOSTERIC REGULATION; BINDING SITES; CATALYSIS; DEUTERIUM; ENZYME ACTIVATION; GLUTAMIC ACID; HYDROGEN-ION CONCENTRATION; KINETICS; MAGNETIC RESONANCE SPECTROSCOPY; MUTAGENESIS, SITE-DIRECTED; PROTONS; PYRUVATE DECARBOXYLASE; PYRUVATES; THERMODYNAMICS; THIAMINE PYROPHOSPHATE; TRANSKETOLASE;

EID: 0031013992     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5296.67     Document Type: Article

References (38)

1. F. J. Alvarez, J. Ermer, G. Hübner, A. Schellenberger, R. L. Schowen, J. Am. Chem. Soc. 113, 8402 (1991).
2. R. Breslow, ibid. 80, 3719 (1958); H. Holzer and K. Beaucamp, Angew. Chem. 71, 776 (1959).
3. R. Breslow, ibid. 80, 3719 (1958); H. Holzer and K. Beaucamp, Angew. Chem. 71, 776 (1959).
4. M. W. Washabaugh and W. P. Jencks, Biochemistry 27, 5044 (1988).
5. J. Crosby and G. E. Lienhard, J. Am. Chem. Soc. 92, 5707 (1970); D. S. Kemp and J. T. O'Brien, ibid., p. 2554.
6. J. Crosby and G. E. Lienhard, J. Am. Chem. Soc. 92, 5707 (1970); D. S. Kemp and J. T. O'Brien, ibid., p. 2554.
7. A. Schellenberger, Angew. Chem. Int. Ed. Engl. 6, 1024 (1967); D. R. Petzold, Stud. Biophys. 54, 159 (1976).
8. A. Schellenberger, Angew. Chem. Int. Ed. Engl. 6, 1024 (1967); D. R. Petzold, Stud. Biophys. 54, 159 (1976).
9. E. J. Crane, J. A. Vaccaro, M. W. Washabaugh, J. Am. Chem. Soc. 115, 8912 (1993); T. K. Harris and M. W. Washabaugh, Biochemistry 34, 13994 (1995); ibid., p. 14001.
10. E. J. Crane, J. A. Vaccaro, M. W. Washabaugh, J. Am. Chem. Soc. 115, 8912 (1993); T. K. Harris and M. W. Washabaugh, Biochemistry 34, 13994 (1995); ibid., p. 14001.
11. E. J. Crane, J. A. Vaccaro, M. W. Washabaugh, J. Am. Chem. Soc. 115, 8912 (1993); T. K. Harris and M. W. Washabaugh, Biochemistry 34, 13994 (1995); ibid., p. 14001.
12. F. Dyda et al., Biochemistry 32, 6165 (1993).
13. Y. Lindqvist, G. Schneider, U. Ermler, M. Sundström, EMBO J. 11, 2373 (1992).
14. M. Nikkola, Y. Lindqvist, G. Schneider, J. Mol. Biol. 238, 387 (1994).
15. Y. A. Muller and G. E. Schulz, Science 259, 965 (1993).
16. Y. A. Muller, G. Schuhmacher, R. Rudolph, G. E. Schulz, J. Mol. Biol. 237, 315 (1994).
17. P. Arjunan et al., ibid. 256, 590 (1996).
18. C. Wikner et al., J. Biol. Chem. 269, 32144 (1994).
19. A. Boiteux and B. Hess, FEBS Lett. 9, 293 (1970).
20. G. Hübner, R. Weidhase, A. Schellenberger, Eur. J. Biochem. 92, 175 (1978).
21. H. Zehender and J. Ullrich, FEBS Lett. 180, 51 (1985).
22. 13C2-ThDP in the samples used for spectra B and C (Fig. 2).
23. 1H-NMR spectra of the supernatant containing only the ThDP were recorded in a 5-mm NMR tube on a Bruker AMX 500-MHz NMR spectrometer.
24. S. Bringer-Meyer, K. L. Schmiz, H. Sahm, Arch. Microbiol. 146, 105 (1986).
25. A. Schellenberger, Chem. Ber. 123, 1489 (1970).
26. R. Golbik et al., Bioorg. Chem. 19, 10 (1991).
27. A. Schellenberger and G. Hübner, Biochem. Educ. 13, 160 (1985); G. Schneider and Y. Lindqvist, Bioorg. Chem. 21, 109 (1993); F. Jordan and Y. H. Mariam, J. Am. Chem. Soc. 100, 2532 (1978); F. Jordan, G. Chen, S. Nishikawa, B. S. Wu, Ann. N. Y. Acad. Sci. 378, 14 (1982); J. Koga, Biochim. Biophys. Acta 1249, 1 (1995).
28. A. Schellenberger and G. Hübner, Biochem. Educ. 13, 160 (1985); G. Schneider and Y. Lindqvist, Bioorg. Chem. 21, 109 (1993); F. Jordan and Y. H. Mariam, J. Am. Chem. Soc. 100, 2532 (1978); F. Jordan, G. Chen, S. Nishikawa, B. S. Wu, Ann. N. Y. Acad. Sci. 378, 14 (1982); J. Koga, Biochim. Biophys. Acta 1249, 1 (1995).
29. A. Schellenberger and G. Hübner, Biochem. Educ. 13, 160 (1985); G. Schneider and Y. Lindqvist, Bioorg. Chem. 21, 109 (1993); F. Jordan and Y. H. Mariam, J. Am. Chem. Soc. 100, 2532 (1978); F. Jordan, G. Chen, S. Nishikawa, B. S. Wu, Ann. N. Y. Acad. Sci. 378, 14 (1982); J. Koga, Biochim. Biophys. Acta 1249, 1 (1995).
30. A. Schellenberger and G. Hübner, Biochem. Educ. 13, 160 (1985); G. Schneider and Y. Lindqvist, Bioorg. Chem. 21, 109 (1993); F. Jordan and Y. H. Mariam, J. Am. Chem. Soc. 100, 2532 (1978); F. Jordan, G. Chen, S. Nishikawa, B. S. Wu, Ann. N. Y. Acad. Sci. 378, 14 (1982); J. Koga, Biochim. Biophys. Acta 1249, 1 (1995).
31. A. Schellenberger and G. Hübner, Biochem. Educ. 13, 160 (1985); G. Schneider and Y. Lindqvist, Bioorg. Chem. 21, 109 (1993); F. Jordan and Y. H. Mariam, J. Am. Chem. Soc. 100, 2532 (1978); F. Jordan, G. Chen, S. Nishikawa, B. S. Wu, Ann. N. Y. Acad. Sci. 378, 14 (1982); J. Koga, Biochim. Biophys. Acta 1249, 1 (1995).
32. 418 → Ala (E418A) TK (13) and 4% for H481A TK (C. Wikner, U. Nilsson, L. Meshalkina, Y. Lindqvist, G. Schneider, in preparation).
33. In addition, a comparatively small increase in the deprotonation rate by pyruvamide activation in the mutant PDC emphasizes that the signal transfer from the regulatory to the active center is probably mediated by E51. The structural events responsible for this step remain to be clarified.
34. H. Neef, K.-D. Kohnert, A. Schellenberger, J. Pract. Chem. 315, 701 (1973).
35. A. Schellenberger, K. Wendler, P. Kreutzberg, G. Hübner, Hoppe-Seyler's Z. Physiol. Chem. 348, 501 (1967).
36. S. König, A. Schellenberger, H. Neef, G. Schneider, J. Biol. Chem. 269, 10879 (1994).
37. First, single-turnover conditions ([enzyme] > [substrate]), which were the basis for their calculations, are not valid because hydrolysis of the activator pyruvamide (1OO mM) increases the initial pyruvate concentration (25 μM) above the enzyme concentration (50 μM). Second, the recombination of ThDP with apo-PDC was not complete within the used time, whereas our experiments were done with the intact holoenzyme after separation of excess free coenzyme. Third, our data explain why an isotope effect for C2-hydrogen exchange could not be observed under the experimental conditions used. Within the recombination time of ThDP with PDC (20 s), the deuterium on C2 of ThDP is completely replaced by a proton (Table 1).
38. We thank D. Wemmer for critical reading of the manuscript. Supported by the Fond der Chemischen Industrie, the Deutsche Forschungsge-meinschaft, the Swedish Natural Science Research Council, and the National Board for Industrial and Technical Development.