1887

Abstract

Protein kinase C (PKC) from mycelium (Basidiomycotina) was purified by affinity chromatography using activated agarose (Affigel 15) coupled with the protein kinase inhibitor -(2-aminoethyl)-5-isoquinolinesulphonamide hydrochloride. Using buffer containing 2·0 M-arginine. PKC activity eluted as a single peak corresponding to a protein with apparent molecular mass of approximately 68000 Da as estimated by SDS-PAGE. The enzyme was not stimulated by Ca alone, and activation by phosphatidylserine (PS) was significant only at 10 M-Ca. However, the activity was enhanced by the addition of 1,2-dioctanoyl--glycerol in the presence of PS and Ca. Additionally, although sensitivity was lower than for mammalian PKC, the enzyme was activated by a tumour-promoting phorbol ester, phorbol-12-myristate 13-acetate (PMA). The saturation concentration of PMA was 50 g ml, which was 5 × 10 times the value for mammalian PKC.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-1-165
1993-01-01
2021-08-05
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/1/mic-139-1-165.html?itemId=/content/journal/micro/10.1099/00221287-139-1-165&mimeType=html&fmt=ahah

References

  1. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principles of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  2. Costagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. 1982; Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. Journal of Biological Chemistry 257:7847–7851
    [Google Scholar]
  3. Eger G. 1968; Investigations on the stimulation of fruit body formation in a Pleurotus from Florida. Theoretical and Applied Genetics 38:23–27
    [Google Scholar]
  4. Eger G. 1976; The action of light and other factors on sporophore initiation in Pleurotus ostreatus.. Mushroom Science IX:575–583
    [Google Scholar]
  5. Fargo A., Nishizuka Y. 1990; Protein kinase C in transmembrane signalling. FEBS Letters 268:350–354
    [Google Scholar]
  6. Favre B., Turian G. 1987; Identification of a calcium- and phospholipid-dependent protein kinase (protein kinase C) in Neurospora crassa. Plant Science 49:15–21
    [Google Scholar]
  7. Hanson B. A. 1991; The effects of lithium on the phosphoinositides and inositol phosphates of Neurospora crassa. Experimental Mycology 15:76–90
    [Google Scholar]
  8. Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. 1984; Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry 23:5036–5041
    [Google Scholar]
  9. Inagaki M., Watanabe M., Hidaka H. 1985; N-(2-aminoethyl)-5-isoquinolinesulfonamide, a newly synthesized protein kinase inhibitor, functions as a ligand in affinity chromatography. Journal of Biological Chemistry 5:2922–2925
    [Google Scholar]
  10. Inagaki M., Yokokura H., Itoh T., Kanmura Y., Kuriyama H., Hidaka H. 1987 Archives of Biochemistry and Biophysics 254:136–141
    [Google Scholar]
  11. Kawamoto S., Hidaka H. 1984; l-(5-isoquinolinesulfonyl)-2-methylpiperazine is a selective inhibitor of protein kinase C in rabbit platelets. Biochemical and Biophysical Research Communications 125:258–264
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  13. Magae Y., Magae J. 1993; Effect of staurosporine on growth and hyphal morphology of source Pleurotus ostreatus. Journal of General Microbiology 139:161–164
    [Google Scholar]
  14. Magae Y., Haga K., Taniguchi H., Sasaki T. 1990; Enzymes of strains of Pleurotus species (basidiomycetes) compared by electrophoresis. Journal of General and Applied Microbiology 36:69–80
    [Google Scholar]
  15. Nishizuka Y. 1986; Studies and prospectives of protein kinase C. Science 233:305–312
    [Google Scholar]
  16. Ogita K., Miyamoto S., Koide H., Iwai T., Oka M., Ando K., Kishimoto A., Ikeda K., Fukami Y., Nishizuka Y. 1990; Protein kinase C in Saccharomyces cerevisiae: comparison with the mammalian enzyme. Proceedings of the National Academy of Sciences of the United States of America 87:5011–5015
    [Google Scholar]
  17. Takagi Y., Katayose Y., Shishido K. 1988; Intracellular levels of cyclic AMP and adenylate cyclase activity during mycelial development in fruiting body formation in Lentinus edodes. FEMS Microbiology Letters 55:275–278
    [Google Scholar]
  18. Takai Y., Kishimoto A., Kikkawa U., Mori T., Nishizuka Y. 1979; Unsaturated diacylglycerol as a possible messenger for the activation of calcium-activated, phospholipid-dependent protein kinase system. Biochemical and Biophysical Research Communications 91:1218–1224
    [Google Scholar]
  19. Yli-Mattila T. 1987; The effect of UV-A light on cAMP level in the basidiomycete Schizophyllum commune. Physiologia Plantarum 69:451–455
    [Google Scholar]
  20. Zadrazil F. 1978; Cultivation of Pleurotus. In The Biology and Cultivation of Edible Mushrooms pp. 521–557 Chang S. T., Hayes W. A. Edited by New York: Academic Press;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-1-165
Loading
/content/journal/micro/10.1099/00221287-139-1-165
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