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. 2000 Mar 1;346(Pt 2):355–358.

Enzyme activity and dynamics: xylanase activity in the absence of fast anharmonic dynamics.

R V Dunn 1, V Réat 1, J Finney 1, M Ferrand 1, J C Smith 1, R M Daniel 1
PMCID: PMC1220860  PMID: 10677353

Abstract

The activity and dynamics of a simple, single subunit enzyme, the xylanase from Thermotoga maritima strain Fj SS3B.1 have been measured under similar conditions, from -70 to +10 degrees C. The internal motions of the enzyme, as evidenced by neutron scattering, undergo a sharp transition within this temperature range; they show no evidence for picosecond-timescale anharmonic behaviour (e.g. local diffusive motions or jumps between alternative conformations) at temperatures below -50 degrees C, whereas these motions are strongly activated at higher temperatures. The activity follows Arrhenius behaviour over the whole of the temperature range investigated, -70 to +10 degrees C. The results indicate that a temperature range exists over which the enzyme rate-limiting step is independent of fast anharmonic dynamics.

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Selected References

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  1. Artymiuk P. J., Blake C. C., Grace D. E., Oatley S. J., Phillips D. C., Sternberg M. J. Crystallographic studies of the dynamic properties of lysozyme. Nature. 1979 Aug 16;280(5723):563–568. doi: 10.1038/280563a0. [DOI] [PubMed] [Google Scholar]
  2. Bauminger E. R., Cohen S. G., Nowik I., Ofer S., Yariv J. Dynamics of heme iron in crystals of metmyoglobin and deoxymyoglobin. Proc Natl Acad Sci U S A. 1983 Feb;80(3):736–740. doi: 10.1073/pnas.80.3.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett W. S., Jr, Steitz T. A. Structure of a complex between yeast hexokinase A and glucose. II. Detailed comparisons of conformation and active site configuration with the native hexokinase B monomer and dimer. J Mol Biol. 1980 Jun 25;140(2):211–230. doi: 10.1016/0022-2836(80)90103-5. [DOI] [PubMed] [Google Scholar]
  4. Cusack S., Doster W. Temperature dependence of the low frequency dynamics of myoglobin. Measurement of the vibrational frequency distribution by inelastic neutron scattering. Biophys J. 1990 Jul;58(1):243–251. doi: 10.1016/S0006-3495(90)82369-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Daniel R. M., Dines M., Petach H. H. The denaturation and degradation of stable enzymes at high temperatures. Biochem J. 1996 Jul 1;317(Pt 1):1–11. doi: 10.1042/bj3170001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daniel R. M., Finney J. L., Réat V., Dunn R., Ferrand M., Smith J. C. Enzyme dynamics and activity: time-scale dependence of dynamical transitions in glutamate dehydrogenase solution. Biophys J. 1999 Oct;77(4):2184–2190. doi: 10.1016/S0006-3495(99)77058-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Daniel R. M., Smith J. C., Ferrand M., Héry S., Dunn R., Finney J. L. Enzyme activity below the dynamical transition at 220 K. Biophys J. 1998 Nov;75(5):2504–2507. doi: 10.1016/S0006-3495(98)77694-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Doster W., Cusack S., Petry W. Dynamical transition of myoglobin revealed by inelastic neutron scattering. Nature. 1989 Feb 23;337(6209):754–756. doi: 10.1038/337754a0. [DOI] [PubMed] [Google Scholar]
  9. Duan Y., Kollman P. A. Pathways to a protein folding intermediate observed in a 1-microsecond simulation in aqueous solution. Science. 1998 Oct 23;282(5389):740–744. doi: 10.1126/science.282.5389.740. [DOI] [PubMed] [Google Scholar]
  10. Ferrand M., Dianoux A. J., Petry W., Zaccaï G. Thermal motions and function of bacteriorhodopsin in purple membranes: effects of temperature and hydration studied by neutron scattering. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9668–9672. doi: 10.1073/pnas.90.20.9668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frauenfelder H., Petsko G. A., Tsernoglou D. Temperature-dependent X-ray diffraction as a probe of protein structural dynamics. Nature. 1979 Aug 16;280(5723):558–563. doi: 10.1038/280558a0. [DOI] [PubMed] [Google Scholar]
  12. Gerstein M., Lesk A. M., Chothia C. Structural mechanisms for domain movements in proteins. Biochemistry. 1994 Jun 7;33(22):6739–6749. doi: 10.1021/bi00188a001. [DOI] [PubMed] [Google Scholar]
  13. Hartmann H., Parak F., Steigemann W., Petsko G. A., Ponzi D. R., Frauenfelder H. Conformational substates in a protein: structure and dynamics of metmyoglobin at 80 K. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4967–4971. doi: 10.1073/pnas.79.16.4967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Huber R., Bennett W. S., Jr Functional significance of flexibility in proteins. Biopolymers. 1983 Jan;22(1):261–279. doi: 10.1002/bip.360220136. [DOI] [PubMed] [Google Scholar]
  15. Jaenicke R. Protein stability and molecular adaptation to extreme conditions. Eur J Biochem. 1991 Dec 18;202(3):715–728. doi: 10.1111/j.1432-1033.1991.tb16426.x. [DOI] [PubMed] [Google Scholar]
  16. Kneller G. R., Smith J. C. Liquid-like side-chain dynamics in myoglobin. J Mol Biol. 1994 Sep 23;242(3):181–185. doi: 10.1006/jmbi.1994.1570. [DOI] [PubMed] [Google Scholar]
  17. More N., Daniel R. M., Petach H. H. The effect of low temperatures on enzyme activity. Biochem J. 1995 Jan 1;305(Pt 1):17–20. doi: 10.1042/bj3050017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Parak F., Frolov E. N., Kononenko A. A., Mössbauer R. L., Goldanskii V. I., Rubin A. B. Evidence for a correlation between the photoinduced electron transfer and dynamic properties of the chromatophore membranes from Rhodospirillum rubrum. FEBS Lett. 1980 Aug 11;117(1):368–372. doi: 10.1016/0014-5793(80)80982-3. [DOI] [PubMed] [Google Scholar]
  19. Parak F., Knapp E. W., Kucheida D. Protein dynamics. Mössbauer spectroscopy on deoxymyoglobin crystals. J Mol Biol. 1982 Oct 15;161(1):177–194. doi: 10.1016/0022-2836(82)90285-6. [DOI] [PubMed] [Google Scholar]
  20. Saul D. J., Williams L. C., Reeves R. A., Gibbs M. D., Bergquist P. L. Sequence and expression of a xylanase gene from the hyperthermophile Thermotoga sp. strain FjSS3-B.1 and characterization of the recombinant enzyme and its activity on kraft pulp. Appl Environ Microbiol. 1995 Nov;61(11):4110–4113. doi: 10.1128/aem.61.11.4110-4113.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Simpson H. D., Haufler U. R., Daniel R. M. An extremely thermostable xylanase from the thermophilic eubacterium Thermotoga. Biochem J. 1991 Jul 15;277(Pt 2):413–417. doi: 10.1042/bj2770413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith J. C. Protein dynamics: comparison of simulations with inelastic neutron scattering experiments. Q Rev Biophys. 1991 Aug;24(3):227–291. doi: 10.1017/s0033583500003723. [DOI] [PubMed] [Google Scholar]
  23. Tilton R. F., Jr, Dewan J. C., Petsko G. A. Effects of temperature on protein structure and dynamics: X-ray crystallographic studies of the protein ribonuclease-A at nine different temperatures from 98 to 320 K. Biochemistry. 1992 Mar 10;31(9):2469–2481. doi: 10.1021/bi00124a006. [DOI] [PubMed] [Google Scholar]
  24. Varley P. G., Pain R. H. Relation between stability, dynamics and enzyme activity in 3-phosphoglycerate kinases from yeast and Thermus thermophilus. J Mol Biol. 1991 Jul 20;220(2):531–538. doi: 10.1016/0022-2836(91)90028-5. [DOI] [PubMed] [Google Scholar]
  25. Wrba A., Schweiger A., Schultes V., Jaenicke R., Závodszky P. Extremely thermostable D-glyceraldehyde-3-phosphate dehydrogenase from the eubacterium Thermotoga maritima. Biochemistry. 1990 Aug 21;29(33):7584–7592. doi: 10.1021/bi00485a007. [DOI] [PubMed] [Google Scholar]
  26. Zechel D. L., Konermann L., Withers S. G., Douglas D. J. Pre-steady state kinetic analysis of an enzymatic reaction monitored by time-resolved electrospray ionization mass spectrometry. Biochemistry. 1998 May 26;37(21):7664–7669. doi: 10.1021/bi980445o. [DOI] [PubMed] [Google Scholar]

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