Abstract
The activity of many proteins in eukaryotic cells is regulated by reversible covalent phosphorylation1. This regulatory modification is often linked to other allosteric controls within the same protein2, and such overlapping regulatory mechanisms are best characterized for glycogen phosphorylase (EC 2.4.1.1). Phosphorylases from different organisms or cell types exhibit markedly contrasting regulatory features3; this makes the enzyme attractive for studying the evolution of interacting molecular regulatory mechanisms4,5. Extensive biochemical and crystallographic studies of rabbit muscle phosphorylase have led to a characterization of five regulatory regions (phosphorylation, glycogen storage, AMP, glucose and purine sites)6–8. Here we report the complete primary structure of the yeast Saccharomyces cerevisiae glycogen phosphorylase, deduced from the sequence of the cloned gene. Regions that are highly conserved between muscle and yeast enzymes include the active site, the glycogen storage site and possibly the glucose and purine inhibition sites. Partial conservation of the residues involved in AMP-binding suggests a binding site for the yeast enzyme inhibitor, glucose 6-phosphate9,10. Other parts of the AMP site and the intersubunit contacts involved in AMP allostery are disrupted in the yeast enzyme by extreme sequence divergence. The poor alignment of amino termini and lack of homology at phosphorylation sites indicate that regulation by reversible phosphorylation evolved independently in yeast and vertebrate phosphorylases.
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References
Cohen, P. Eur. J. Biochem. 151, 439–448 (1985).
Cohen, P. Control of Enzyme Activity 2nd edn, 1–96 (Chapman & Hall, London, 1983).
Graves, D .J. & Wang, T. H. in The Enzymes vol. 7, 3rd edn (ed. Boyer, P.), 430–482 (Academic, London, 1982).
Fischer, E. H., Pocker, A. & Saari, J. C. Essays Biochem. 6, 23–68 (1970).
Palm, D., Goerl, R. & Burger, K. J. Nature 313, 500–502 (1985).
Fletterick, R. J., & Sprang, S. R. Acc. chem. Res., 15, 361–369 (1983).
Fletterick, R. J., & Madsen, N. B. A. Rev. Biochem., 49, 831–861 (1980).
Dombradi, V., Int. J. Biochem. 13, 125–139. 1981).
Fosset, M., Muir, L. W., Nielsen, L. D. & Fischer, E. H. Biochemistry 10, 4105–4113 (1971).
Sagardia, F., Gotay, I. & Rodriquez, M. Biochem. biophys. Res. Comm. 42, 829–935 (1971).
Kasvinsky, P. J., Schechosky, S. & Fletterick, R. J. J. biol. Chem., 253, 9102–9106 (1978).
Guenard, D., Morange, M. & Buc, H. Eur. J. Biochem. 76, 447–452 (1977).
Stura, E. A. et al. J. molec. Biol. 170, 529–565 (1983).
Sprang, S. et al. Biochemistry 21, 2036–2048 (1982).
Sprang, S. R. & Fletterick, R. J. J. molec. Biol. 131, 523–551 (1979).
Lorek, A. et al. Biochem. J. 218, 45–60 1984).
Titani, K. et al. Proc. natn. Acad. Sci. U.S.A. 74, 4762–4766 (1977).
Lerch, K., & Fischer, E. H. Biochemistry 14, 2009–2014 (1975).
Becker, J.-U. Wingender-Drissen, R. & Schiltz, E. Archs Biochem. Biophys. 225, 667–678 (1983).
Pohlig, G. Wingender-Drissen, R. & Becker, J.-U. Biochem. biophys. Res. Commun. 114, 331–338 (1983).
Cohen, P., Saari, J. C. & Fischer, E. H. Biochemistry 12, 5233–5241 (1973).
Schiltz, E., Palm, D. & Klein, H. W. FEBS Lett. 109, 59–62. (1980).
Lacks, S. A., Dunn, J. J. & Greenberg, B. Cell 31, 327–336 (1982).
Nakano, K., Fukui, T., & Matsubara, H. J. biol. Chem. 255, 9255–9261 (1980).
Newgard, C. B., Nakano, K., Hwang, P. K. & Fletterick, R. J. Proc. natn. Acad. Sci. U.S.A. (in the press).
Sprang, S., & Fletterick, R. J. Biophys. J. 32, 175–192 (1980).
Madsen, N. B. & Scheckosky, S. J. biol. Chem., 242, 3301–3307 (1967).
Wingender-Drissen, R. & Becker, J.-U. Biochim. biophys. Acta 743, 343–350 (1983).
Matsumoto, K., Uno, I., Kato, K. & Ishikawa, T. Yeast 1, 25–38 1985).
Carlson, M., & Botstein, D. Cell 28, 145–154, (1982).
Southern, E. J. molec. Biol. 98, 503–517, 1975).
Sanger, F., Coulson, A. R., Barrell, B. G., Smith, A. J. H. & Roe, B. A. J. molec. Biol. 143, 161–178, (1980).
Nakano, K., Hwang, P. K. & Fletterick, R. J. FEBS Lett. (in the press).
Dayhoff, M. O. (ed.) in Atlas of Protein Sequence and Structure Vol. 5, Suppl. 2, 3–8, (Natn. Biomed. Res. Fdn, Silver Spring, Maryland, 1976).
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Hwang, P., Fletterick, R. Convergent and divergent evolution of regulatory sites in eukaryotic phosphorylases. Nature 324, 80–84 (1986). https://doi.org/10.1038/324080a0
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DOI: https://doi.org/10.1038/324080a0
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