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Abstract

  1. Top of page
  2. Abstract
  3. REFERENCES

The in vivo phosphorylation state of glycogen synthase was re-examined by fast-atom-bombardment mass spectrometry and a procedure in which phosphoserine residues are first converted to S-ethylcysteine. In animals injected with the β-adrenergic antagonist propranolol, the phosphorylation sites in the N-terminal (N) and C-terminal (C) cyanogen bromide peptides were identified as the serine residues at N7, the region C28-C39, C42, C46 and C100. In animals injected with adrenalin, the phosphorylation of N7 increased from 0.6 to 0.8 mol/mol, the region C28-C39 from 0.7 to 1.2 mol/mol and C100 from 0.3 to 0.6 mol/mol. The phosphorylation states of C42 (0.7 mol/mol) and C46 (0.9 mol/mol) were unchanged. In addition, two further serine residues became phosphorylated at positions N10 (0.5 mol/mol) and C87 (0.5 mol/mol), which were not phosphorylated in the absence of adrenalin.

Residues N10 and C42 have not been recognized as in vivo sites of phosphorylation previously. The results suggest that N10 is phosphorylated by a novel protein kinase which may be activated by cyclic-AMP-dependent protein kinase. The phosphorylation of C42 is likely to be catalysed by glycogen synthase kinase 3. The protein kinases responsible for phosphorylating N7, the region C28–C39, C46, C87 and C100 in vivo and the molecular mechanisms by which adrenalin inactivates glycogen synthase in vivo are discussed.

Residue N3, a major site phosphorylated by casein kinase-I in vitro is not phosphorylated in vivo. This and other evidence indicates that casein kinase-I is not a glycogen synthase kinase in vivo.

Abbreviations
FAB MS

fast-atom-bombardment mass spectrometry

Pth

phenylthiohydantoin

EtCys

S-ethylcysteine

Enzymes
 

Glycogen synthase (EC 2.4.1.11)

 

protein kinase (EC 2.7.1.37)

 

protein phosphatase (EC 3.1.3.16)

 

trypsin (EC 3.4.21.4)

 

chymotrypsin (EC 3.4.21.1)

 

Staphylococcus aureus V8 proteinase (EC 3.4.21.19)

REFERENCES

  1. Top of page
  2. Abstract
  3. REFERENCES
  • 1
    Picton, C., Aitken, A., Bilham, T. & Cohen, P. (1982) Eur. J. Biochem. 124, 3745.
  • 2
    Cohen, P., Holmes, C. F. B., Poulter, L., Gibson, B. & Williams, D. H. (1986) Biochem. Biophys. Res. Commun. 137, 5425545.
  • 3
    Payne, M. E., Schworer, C. M. & Soderling, T. R. (1983) J. Biol. Chem. 258, 23762382.
  • 4
    Woodgett, J. R., Davison, M. T. & Cohen, P. (1983) Eur. J. Biochem. 136, 481487.
  • 5
    McGuinness, T. L., Lai, Y., Greengard, P., Woodgett, J. R. & Cohen, P. (1983) FEBS Lett. 163, 329334.
  • 6
    Embi, N., Rylatt, D. B. & Cohen, P. (1979) Eur. J. Biochem. 100, 339347.
  • 7
    Soderling, T. R., Sheorain, V. S. & Ericsson, L. H. (1979) FEBS Lett. 106, 181184.
  • 8
    Embi, N., Parker, P. J. & Cohen, P. (1981) Eur. J. Biochem. 115, 405413.
  • 9
    Cohen, P., Yellowlees, D., Aitken, A., Donella-Deana, A., Hemmings, B. A. & Parker, P. J. (1982) Eur. J. Biochem. 124, 2135.
  • 10
    Rylatt, D. B., Aitken, A., Bilham, T., Condon, G. D., Embi, N. & Cohen, P. (1980) Eur. J. Biochem. 107, 529537.
  • 11
    Cohen, P. (1986) The Enzymes (3rd edn) (Boyer, P. D. & Krebs, E. G. eds) vol. 17, pp. 461497, Academic Press Orlando , FL .
  • 12
    Parker, P. J., Embi, N., Caudwell, F. B. & Cohen, P. (1982) Eur. J. Biochem. 124, 4755.
  • 13
    Paarker, P. J., Caudwell, F. B. & Cohen, P. (1983) Eur. J. Biochem. 130, 227234.
  • 14
    Kuret, J., Woodgett, J. R. & Cohen, P. (1985) Eur. J. Biochem. 151, 3948.
  • 15
    Cohen, P., Watson, D. C. & Dixon, G. H. (1975) Eur. J. Biochem. 51, 7992.
  • 16
    Yeaman, S. J. & Cohen, P. (1975) Eur. J. Biochem. 51, 93104.
  • 17
    Proud, C. G., Rylatt, D. B., Yeaman, S. J. & Cohen, P. (1977) FEBS Lett. 80, 435442.
  • 18
    Sheorain, V. S., Corbin, J. D. & Soderling, T. R. (1985) J. Biol. Chem. 260, 15671572.
  • 19
    Ahmad, Z., DePaoli-Roach, A. A. & Roach, P. J. (1985) FEBS Lett. 179, 96100.
  • 20
    Ahmad, Z., Lee, F. T., DePaoli-Roach, A. A. & Roach, P. J. (1986) Arch. Biochem. Biophys. 250, 329335.
  • 21
    Meyer, H. E., Hoffmann-Posorske, E., Korte, H. & Heilmeyer, L. M. G. (1986) FEBS Lett. 204, 6166.
  • 22
    Holmes, C. F. B. (1987) FEBS Lett. 215, 2124.
  • 23
    Gibson, B. W., Falick, A. M., Burlingame, A. L., Kenyon, G. L., Poulter, L., Williams, D. H. & Cohen, P. (1987) in 6th International conference on methods in sequence analysis (Walsh, K. ed.) pp. 463478, Humana Press, Clifton , NJ .
  • 24
    Nimmo, H. G., Proud, C. G. & Cohen, P. (1976) Eur. J. Biochem. 68, 2130.
  • 25
    Caudwell, F. B., Antoniw, J. F. & Cohen, P. (1978) Eur. J. Biochem. 86, 511518.
  • 26
    Poulter, L., Ang, S. G., Williams, D. H. & Cohen, P. (1987) Biochim. Biophys. Acta 929, 296301.
  • 27
    Pitcher, J., Smythe, C., Campbell, D. G. & Cohen, P. (1987) Eur. J. Biochem. 169, 497502.
  • 28
    Huang, T. S. & Krebs, E. G. (1979) FEBS Lett. 98, 6670.
  • 29
    Holmes, C. F. B., Campbell, D. G., Caudwell, F. B., Aitken, A. & Cohen, P. (1986) Eur. J. Biochem. 155, 173182.
  • 30
    Tarr, G. E. (1982) 4th International conference on methods in sequence analysis (Elzinga, M. ed.) p. 223, Humana Press, Clifton , NJ .
  • 31
    Naylor, S., Findeis, A. F., Gibson, B. W. & Williams, D. H. (1986) J. Am. Chem. Soc. 108, 63596363.
  • 32
    Sheorain, V. S., Juhl, H., Bass, M. & Soderling, J. R. (1984) J. Biol. Chem. 259, 70247030.
  • 33
    Huang, K. P., Singh, T. J., Akatsuka, A., Shapiro, S. G., Vandenheede, J. R. & Merlevede, W. (1984) Arch. Biochem. Biophys. 232, 111117.
  • 34
    Ahmad, Z., Camici, M., DePaoli-Roach, A. A. & Roach, P. J (1984) J. Biol. Chem. 259, 34203428.
  • 35
    Woodgett, J. R. (1984) Ph. D. Thesis, University of Dundee.
  • 36
    Fiol, C. J., Wang, Y., Mahrenholz, A., Roeske, R. & Roach, P. J. (1987) J. Biol. Chem. 262, 1404214048.
  • 37
    Picton, C., Woodgett, J. R., Hemmings, B. A. & Cohen, P. (1982) FEBS Lett. 150, 191196.
  • 38
    De Paoli-Roach, A. A., Ahmed, Z., Camici, M., Lawrence, J. C. & Roach, P. J. (1983) J. Biol. Chem. 258, 1070210709.
  • 39
    Hemmings, B. A., Yellowlees, D., Kernohan, J. C. & Cohen, P. (1981) Eur. J. Biochem. 119, 443451.
  • 40
    Stralfors, P., Hiraga, A. & Cohen, P. (1985) Eur. J. Biochem. 149, 295303.
  • 41
    Foulkes, J. G. & Cohen, P. (1979) Eur. J. Biochem. 97, 251256.
  • 42
    Hiraga, A. & Cohen, P. (1987) Eur. J. Biochem. 161, 763796.
  • 43
    Reopstorff, R. & Fahlman, J. (1984) Biomed. Mass Spectrom. 11, 601.