Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 Jul 1;349(Pt 1):211–215. doi: 10.1042/0264-6021:3490211

Investigation of the slow inhibition of almond beta-glucosidase and yeast isomaltase by 1-azasugar inhibitors: evidence for the 'direct binding' model.

A Lohse 1, T Hardlei 1, A Jensen 1, I W Plesner 1, M Bols 1
PMCID: PMC1221139  PMID: 10861230

Abstract

(-)-1-Azafagomine [(3R,4R,5R)-4,5-dihydroxy-3-hydroxymethylhexahydropyridazine; inhibitor 1] is a potent glycosidase inhibitor designed to mimic the transition state of a substrate undergoing glycoside cleavage. The inhibition of glycosidases by inhbitor 1 and analogues has been found to be a relatively slow process. This 'slow inhibition' process was investigated in the inhibition of almond beta-glucosidase and yeast isomaltase by inhibitor 1 and analogues. Progress-curve experiments established that the time-dependent inhibition of both enzymes by inhibitor 1 was a consequence of relatively slow dissociation and association of the inhibitor from and to the enzyme, and not a result of slow interchanges between protein conformations. A number of hydrazine-containing analogues of inhibitor 1 also inhibited beta-glucosidase and isomaltase slowly, while the amine isofagomine [(3R,4R,5R)-3,4-dihydroxy-5-hydroxymethylpiperidine; inhibitor 5] only inhibited beta-glucosidase slowly. Inhibitor 1 and related inhibitors were found to leave almond beta-glucosidase with almost identical rate constants, so that the difference in K(i) values depended almost entirely on changes in the binding rate constant, k(on). The same trend was observed for the inhibition of yeast isomaltase by inhibitor 1 and a related inhibitor. The values of the rate constants were obtained at 25 degrees C and at pH 6.8.

Full Text

The Full Text of this article is available as a PDF (128.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bernacki R. J., Niedbala M. J., Korytnyk W. Glycosidases in cancer and invasion. Cancer Metastasis Rev. 1985;4(1):81–101. doi: 10.1007/BF00047738. [DOI] [PubMed] [Google Scholar]
  2. Dale M. P., Ensley H. E., Kern K., Sastry K. A., Byers L. D. Reversible inhibitors of beta-glucosidase. Biochemistry. 1985 Jul 2;24(14):3530–3539. doi: 10.1021/bi00335a022. [DOI] [PubMed] [Google Scholar]
  3. Dennis J. W., Laferté S., Waghorne C., Breitman M. L., Kerbel R. S. Beta 1-6 branching of Asn-linked oligosaccharides is directly associated with metastasis. Science. 1987 May 1;236(4801):582–585. doi: 10.1126/science.2953071. [DOI] [PubMed] [Google Scholar]
  4. Ernholt B. V., Thomsen Ib B., Lohse A., Plesner I. W., Jensen K. B., Hazell R. G., Liang X., Jakobsen A., Bols M. Enantiospecific synthesis of 1-azafagomine. Chemistry. 2000 Jan;6(2):278–287. doi: 10.1002/(SICI)1521-3765(20000117)6:2<278::AID-CHEM278>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  5. Grover A. K., Macmurchie D. D., Cushley R. J. Studies on almond emulsin beta-D-glucosidase. I. Isolation and characterization of a bifunctional isozyme. Biochim Biophys Acta. 1977 May 12;482(1):98–108. doi: 10.1016/0005-2744(77)90358-8. [DOI] [PubMed] [Google Scholar]
  6. Hanozet G., Pircher H. P., Vanni P., Oesch B., Semenza G. An example of enzyme hysteresis. The slow and tight interaction of some fully competitive inhibitors with small intestinal sucrase. J Biol Chem. 1981 Apr 25;256(8):3703–3711. [PubMed] [Google Scholar]
  7. Karpas A., Fleet G. W., Dwek R. A., Petursson S., Namgoong S. K., Ramsden N. G., Jacob G. S., Rademacher T. W. Aminosugar derivatives as potential anti-human immunodeficiency virus agents. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9229–9233. doi: 10.1073/pnas.85.23.9229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kati W. M., Saldivar A. S., Mohamadi F., Sham H. L., Laver W. G., Kohlbrenner W. E. GS4071 is a slow-binding inhibitor of influenza neuraminidase from both A and B strains. Biochem Biophys Res Commun. 1998 Mar 17;244(2):408–413. doi: 10.1006/bbrc.1998.8282. [DOI] [PubMed] [Google Scholar]
  9. Kleinschmidt T., Glossmann H., Horst J. Zur Kenntnis des Süssmandel-Emulsins. Weitere Charakterisierung der Komponenten A und B. Hoppe Seylers Z Physiol Chem. 1970 Mar;351(3):349–358. [PubMed] [Google Scholar]
  10. Kobata A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem. 1979 Nov 15;100(1):1–14. doi: 10.1016/0003-2697(79)90102-7. [DOI] [PubMed] [Google Scholar]
  11. Kornfeld R., Kornfeld S. Comparative aspects of glycoprotein structure. Annu Rev Biochem. 1976;45:217–237. doi: 10.1146/annurev.bi.45.070176.001245. [DOI] [PubMed] [Google Scholar]
  12. Legler G. Glycoside hydrolases: mechanistic information from studies with reversible and irreversible inhibitors. Adv Carbohydr Chem Biochem. 1990;48:319–384. doi: 10.1016/s0065-2318(08)60034-7. [DOI] [PubMed] [Google Scholar]
  13. Marshall J. J. Application of enzymic methods to the structural analysis of polysaccharides: part I. Adv Carbohydr Chem Biochem. 1974;30:257–370. doi: 10.1016/s0065-2318(08)60267-x. [DOI] [PubMed] [Google Scholar]
  14. Merkler D. J., Brenowitz M., Schramm V. L. The rate constant describing slow-onset inhibition of yeast AMP deaminase by coformycin analogues is independent of inhibitor structure. Biochemistry. 1990 Sep 11;29(36):8358–8364. doi: 10.1021/bi00488a023. [DOI] [PubMed] [Google Scholar]
  15. Morrison J. F., Walsh C. T. The behavior and significance of slow-binding enzyme inhibitors. Adv Enzymol Relat Areas Mol Biol. 1988;61:201–301. doi: 10.1002/9780470123072.ch5. [DOI] [PubMed] [Google Scholar]
  16. Pegg M. S., von Itzstein M. Slow-binding inhibition of sialidase from influenza virus. Biochem Mol Biol Int. 1994 Apr;32(5):851–858. [PubMed] [Google Scholar]
  17. Ploux O., Breyne O., Carillon S., Marquet A. Slow-binding and competitive inhibition of 8-amino-7-oxopelargonate synthase, a pyridoxal-5'-phosphate-dependent enzyme involved in biotin biosynthesis, by substrate and intermediate analogs. Kinetic and binding studies. Eur J Biochem. 1999 Jan;259(1-2):63–70. doi: 10.1046/j.1432-1327.1999.00006.x. [DOI] [PubMed] [Google Scholar]
  18. Robinson K. M., Begovic M. E., Rhinehart B. L., Heineke E. W., Ducep J. B., Kastner P. R., Marshall F. N., Danzin C. New potent alpha-glucohydrolase inhibitor MDL 73945 with long duration of action in rats. Diabetes. 1991 Jul;40(7):825–830. doi: 10.2337/diab.40.7.825. [DOI] [PubMed] [Google Scholar]
  19. Yiallouros I., Vassiliou S., Yiotakis A., Zwilling R., Stöcker W., Dive V. Phosphinic peptides, the first potent inhibitors of astacin, behave as extremely slow-binding inhibitors. Biochem J. 1998 Apr 15;331(Pt 2):375–379. doi: 10.1042/bj3310375. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES