Advertisement

Science in China Series C: Life Sciences

, Volume 41, Issue 3, pp 245–250 | Cite as

Activation and conformational changes of adenylate kinase in urea solution

  • Hongjie Zhang
  • Xianming Pan
  • Junmei Zhou
  • Kihara Hiroshi
Article

Abstract

The activation and inactivation of adenylate kinase during deneturation in urea are compared with changes in UV absorbance at 287 nm. CD spectrum change at 222 nm, fluorescence intensity of ANS biding and small angle of X-ray scattering. At 1 mol/L, of urea the enzyme is activated 1.5-fold companied with a subtie decressing of its second structure, whereas its tertiary structure is fairly resistant to denaturation. By comparing the studies of the crystal structure and the mechanism of the catalysis of adenylste kinase, the activation is believed to result the effect that low concentration of urea increases the flexibility of the active site of the enzyme. This suggestion was confirmed by the results of the fluorescence intensity changes of ANS binding to adenylate kinase versus the concentration of urea.

Key words

adenylate kinase urea denaturation activation flexibillity of aetive site 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Zhang, Y. L., Zhou, J. M., Tsou, C. L., Inactivation precedes conformation change during thermal denaturation of adenylate kinase,Biochim. Biophys. Acta, 1993, 1164: 61PubMedGoogle Scholar
  2. 2.
    Zhang, Y. L., Zhou, J. M., Tsou, C. L., Sequential unfolding of adenylate kinase during denaturation by guanidine hydrochloride,Biochim. Biophys. Acta, 1996, 1295: 239.PubMedGoogle Scholar
  3. 3.
    Tsou, C. L., Inactivation precedes overall molecular conformation changes during enzyme denaturation.Biochim. Biophys. Acta, 1995, 1253: 151.PubMedGoogle Scholar
  4. 4.
    Glatter, O., Kratky, O.,Small-Angle X-ray Scattering, New York: Academic Press. 1982, 119.Google Scholar
  5. 5.
    Yao, Q. Z., Tian, M., Tsou, C. L., Comparison of the rates of inactivation and conformational changes of creatine kinase during urea denaturation,Biochemistry, 1984, 23: 2740.PubMedCrossRefGoogle Scholar
  6. 6.
    Liu, W., Tsou, C. L., Activity change during unfolding of bovine pancreatic ribonuclease A in guenidine hydrochloride,Biochim. Biophys. Acta. 1987, 916: 455.PubMedGoogle Scholar
  7. 7.
    Xie, G. F., Tsou, C. L., Conformational and activity changes during guanidine deneturation of D-glyceraldehyde-3-phosphate dehydrogenase,Biochim, Biophys. Acta. 1987, 911: 19.Google Scholar
  8. 8.
    Tsou, C. L., Conformational flexibility of enzyme active sites,Science, 1993, 262: 380.PubMedCrossRefGoogle Scholar
  9. 9.
    Gerstein, M., Schulz, G., Chorhia, C., Domain closure in adenylate kinase, joints on either side of two helices close like neighboring fingers,J. Mol. Biol., 1993, 229: 494.PubMedCrossRefGoogle Scholar
  10. 10.
    Hamada, M., Kuby, S. A., Studies on adenosine triphospthate transphosphorylase XIII—kinetic properties of the crystalline rabbit muscle ATP-AMP transphosphorylase (adenylate kinase) and a comparison with the crystalline calf muscle and liver adenylate kinase,Arch. Biochem, Biophy., 1978, 190: 772.CrossRefGoogle Scholar
  11. 11.
    Semistnov, G. V., Rdionova, N. A., Razgulyaev, O. I. et al., Study of the “molten globule” intermediate state in protein folding by a hydrophobic fluorescent probe,Biopolymers, 1991, 31: 119.CrossRefGoogle Scholar
  12. 12.
    Creighton, T. E.,Protein Folding, New York: W. H. Freeman, 1992, 243–300.Google Scholar
  13. 13.
    Pai, E. F., Sachsenheimer, W., Schirmer, R. H. et al., Substrate positions and induced-fit in cryatalline adenylate kinese.J. Mol. Biol., 1977, 114: 37.PubMedCrossRefGoogle Scholar
  14. 14.
    Muller, C. W., Schulz, G. E., Structure ol the complex between adenylate kinase fromEsrherichia coli and the inhibitor Ap5A refined at 1.9 Å resolution,J. Mol. Biol., 1992, 224: 159.PubMedCrossRefGoogle Scholar
  15. 15.
    Fan, Y. X., Ju, M., Zhou, J. M. et al., Activation nf chicken liver dihydrofolate reductase in concentrated urea solutions.Biochim. Biophs. Acta, 1995, 1252: 151.Google Scholar

Copyright information

© Science in China Press 1998

Authors and Affiliations

  • Hongjie Zhang
    • 1
  • Xianming Pan
    • 1
  • Junmei Zhou
    • 1
  • Kihara Hiroshi
    • 2
  1. 1.State Key Laboratory of Biomacromolecule, Insritute of BiophysicsChinese Academy of SciencesBeijingChina
  2. 2.Physics LaboratoryKansai Medical UniversityOaskaJapan

Personalised recommendations