1887

Abstract

1,3--Glucan synthase activity has been detected in a membrane fraction extracted from the mycelium of the filamentous fungus The enzyme was solubilized by CHAPS and stabilized by filtration on a Bio-gel P30 column. Highest activity was obtained in the early exponential phase of growth. Four factors - GTP, NaF, sucrose and EDTA - added during the extraction procedure, were essential for optimal 1,3--glucan synthase activity. The soluble enzyme preparation was photolabelled with 5-azido-[P]UDP-glucose and 5-IASA-UDP-glucose which bind covalently to the enzyme after UV irradiation. These UDP-glucose substrate analogues were competitive inhibitors of the enzyme with a of 1·42 m and 0·3 m for 5-azido-UDP-glucose and 5-ASA-UDP-glucose, respectively ( for UDP-glucose = 1·9 m). Potential UDP-glucose-binding polypeptides were identified with molecular masses of 31, 50 and 115 kDa.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-12-3071
1993-12-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/12/mic-139-12-3071.html?itemId=/content/journal/micro/10.1099/00221287-139-12-3071&mimeType=html&fmt=ahah

References

  1. Awald P., Zugel M., Monks C., Frost D., Selitrennikoff C.P. 1993; Purification of 1,3-β-glucan synthase from Neurospora crassa by product entrapment. Experimental Mycology 17:130–141
    [Google Scholar]
  2. Beauvais A., Latgé J. P. 1989; Chitin and β(1-3) glucan synthases in protoplastic Entomophthorales. Archives of Microbiology 152:229–236
    [Google Scholar]
  3. Bradford M.M. 1976; A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  4. Bulone V., Girard V., Févre M. 1990; Separation and partial purification of 1,3-β-glucan and 1,4-βglucan synthases from Sapro- legnia. Plant Physiology 94:1748–1755
    [Google Scholar]
  5. Cabib E., Kang M.S. 1987; Fungal 1,3-β-glucan synthase. Methods in Enzymology 138:637–642
    [Google Scholar]
  6. Cerenius L., Söderhäll K. 1984; Isolation and properties of β glucan synthetase from the aquatic fungus, Aphanomyces astaci. Physiologia Plantarum 60:247–252
    [Google Scholar]
  7. Delmer D.P., Solomon M., Read S.M. 1991; Direct photolabeling with [32P]UDP-glucose for identification of a subunit of cotton fiber Callose synthase. Plant Physiology 95:556–563
    [Google Scholar]
  8. Dhugga K.S., Ray P. M. 1991; A 55 kDa plasma membraneassociated polypeptide is involved in p- 1,3-glucan synthase activity in pea tissue. FEBS Letters 278:283–286
    [Google Scholar]
  9. Drake R.R., Evans R.K, Haley B.E. 1989; Synthesis and properties of 5-azido-UDP-glucose. Journal of Biological Chemistry 264:11928–11933
    [Google Scholar]
  10. Fèvre M. 1979; Digitonin solubilization and protease stimulation of β glucan synthetases of Saprolegnia. Zeitschrift für Pflanzen- physiologie 95-S:129–140
    [Google Scholar]
  11. Fink J., Jeblick W., Kauss H. 1990; Partial purification and immunological characterization of 1,3-βglucan synthase from suspension cells of Glycine max. Planta 181:343–348
    [Google Scholar]
  12. Frost D.J., Read S.M., Drake R.R., Haley B.E., Wasserman B.P. 1990; Identification of the UDP-glucose-binding polypeptide of callose synthase from Beta vulgaris L. by photoaffinity labeling with 5-azido-UDP-glucose. Journal of Biological Chemistry 265:2162–2167
    [Google Scholar]
  13. Frost D.J., Drake R.R., Wasserman B.P. 1992; (l,3)-β-Glucan synthase from Saccharomyces cerevisiae: in vitro activation by β lactoglobulin or Brij-25, and photoaffinity labeling of enriched microsomal fractions with 5-azido-UDP-Glc and 8-azido-GTP. Current Microbiology 24:295–300
    [Google Scholar]
  14. Hayashi T., Read S.M., Bussel J., Thelen M., Lin F.-C., Brown R.M., Delmer D.P. 1987; UDP-glucose:(l-3)-βglucan synthases from mung bean and cotton. Plant Physiology 83:1054–1062
    [Google Scholar]
  15. Kamat U., Garg R., Sharma C.B. 1992; Purification to homogeneity and characterization of a 1,3-β-glucan (callose) synthase from germinating Arachis hypogaea cotyledons. Archives of Biochemistry and Biophysics 298:731–739
    [Google Scholar]
  16. Kang M.S., Cabib E. 1986; Regulation of fungal cell wall growth: a guanine nucleotide-binding, proteinaceous component required for activity of (l-3)-βd-glucan synthase. Proceedings of the National Academy of Sciences of the United States of America 835808–5812
    [Google Scholar]
  17. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  18. Larriba G., Andaluz E. 1991; Biosynthesis of yeast glucan. In Fungal Cell Wall and Immune Response pp. 49–68 Edited by Latge J. P., Boucias D. Heidelberg: Springer-Verlag;
    [Google Scholar]
  19. Larriba G., Morales M., Ruiz-Herrera J. 1981; Biosynthesis of β-glucan microfibrils by cell-free extracts from Saccharomyces cerevisiae. Journal of General Microbiology 124:375–383
    [Google Scholar]
  20. Lawson S.G., Sabin R.D., Sloan M.E., Drake R.R., Haley B.E., Wasserman B.P. 1989; UDP-glucose:(l- 3)-β-glucan synthase from Daucus carota L. Plant Physiology 90:101–108
    [Google Scholar]
  21. Lin F.C., Brown R.M., Drake R.R., Haley B.E. 1990; Identification of the uridine 5´-diphosphoglucose (UDP-Glc) binding subunit of cellulose synthase in Acetobacter xylinum using the photo affinity probe 5-azido-UDP-Glc. Journal of Biological Chemistry 265:4782–4784
    [Google Scholar]
  22. Meikle P.J., Ng K.F., Johnson E., Hoogenraad N.T., Stone B.A. 1991; The β-glucan synthase from Lolium multiflorum. Journal of Biological Chemistry 266:22569–22581
    [Google Scholar]
  23. Montreuil J., Bouquelet S., Debray H., Fournet B., Spik G., Strecker G. 1986; Glycoprotein. In Carbohydrate Analysis, a Practical Approach pp. 143–204 Edited by Champlin M. F., Kennedy J. F. Oxford: IRL Press;
    [Google Scholar]
  24. Ohana P, Delmer D. P., Volman G., Steffens J.C., Matthews D.E., Benziman M. 1992; βFurfuryl-βglucoside: an endogenous activator of higher plant UDP-glucose:(l-3)-βglucan synthase. Plant Physiology 98:708–715
    [Google Scholar]
  25. Orlean P.A.B., Ward S.M. 1983; Sodium fluoride stimulates (l-3)-β-d-glucan synthase from Candida albicans. FEMS Microbiology Letters 18:31–35
    [Google Scholar]
  26. Quigley D.R., Hrmova M., Selitrennikoff P. 1988; β(l-3) Glucan synthase of Neurospora crassa: solubilization and partial characterization. Experimental Mycology 12:141–150
    [Google Scholar]
  27. Radominska A., Drake R.R. 1993; Synthesis and uses of azido- nucleoside diphosphate sugar photoaffinity analogs. Methods in Enzymology 230: in the Press
    [Google Scholar]
  28. Szaniszlo P.J., Kang M.S., Cabib E. 1985; Stimulation of β(l-3) glucan synthetase of various fungi by nucleoside triphosphates: generalized regulatory mechanism for cell wall biosynthesis. Journal of Bacteriology 161:118–1194
    [Google Scholar]
  29. Wessels J.G.H. 1986; Cell wall synthesis in apical growth. International Review of Cytology 104:37–97
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-12-3071
Loading
/content/journal/micro/10.1099/00221287-139-12-3071
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error