Note. Residues in glycogen synthase are numbered by their distance from the start of the N-terminal cyanogen bromide fragment, CB-N (N7, N10), and the C-terminal cyanogen bromide fragment, CB-C (C30, C38, C42, C46, C87, C100). See Fig. 1.
The molecular mechanism by which adrenalin inhibits glycogen synthesis
Article first published online: 3 MAR 2005
European Journal of Biochemistry
Volume 199, Issue 3, pages 713–722, August 1991
How to Cite
NAKIELNY, S., CAMPBELL, D. G. and COHEN, P. (1991), The molecular mechanism by which adrenalin inhibits glycogen synthesis. European Journal of Biochemistry, 199: 713–722. doi: 10.1111/j.1432-1033.1991.tb16175.x
- Issue published online: 3 MAR 2005
- Article first published online: 3 MAR 2005
- (Received January 21/March 3, 1991) – EJB 91 0112
Improved methodology was used to establish that the phosphorylation of a serine located 10 residues from the N-terminus of glycogen synthase (N10) increases from 0.12 mol · mol−1 to 0.54 mol · mol−1in vivo in response to adrenalin. The only ‘N10 kinase’ detected in muscle extracts was casein kinase-1 (CK1), although its activity was unaffected by injection of adrenalin in vivo or by incubation with cyclic-AMP-dependent protein kinase and MgATP in vitro. Prior phosphorylation of the serine residue N7 by phosphorylase kinase increased sixfold the rate of phosphorylation of glycogen synthase by CK1, and altered the specificity of CK1 so that it phosphorylated the serine residue N10 specifically. Stoichiometric phosphorylation of N7 decreased the activity ratio (∓ glucose 6-phosphate) of glycogen synthase from 0.80 to 0.45, and subsequent phosphorylation of N10 to 0.8 mol · mol−1 produced a further decrease to 0.17, demonstrating that N10 phosphorylation inhibits glycogen synthase. The major ‘N10 phosphatase’ in skeletal muscle extracts was identified as the glycogen-associated form of protein phosphatase-1 (PP1G), accounting for approximately 75% of the N10 phosphatase activity in the extracts and about 90% of the activity in isolated glycogen particles. Phosphorylation of N10, after prior phosphorylation of N7, decreased the rate of dephosphorylation of N7. These results, in conjunction with previous findings, establish that adrenalin inhibits glycogen synthase by increasing the phosphorylation of N7, N10 and three further serines located 30, 34 and 38 residues from the start of the C-terminal CNBr peptide (termed the region C30–C38). They also indicate that increased phosphorylation of N10, the region C30–C38, and perhaps N7, is initiated through the inhibition of PP1G by adrenalin, which results from phosphorylation of its glycogen-targetting subunit by cyclic-AMP-dependent protein kinase [Hubbard, M. J. & Cohen, P. (1989) Eur. J. Biochem. 186, 711–716]. The conclusion that direct phosphorylation of glycogen synthase by cyclic-AMP-dependent protein kinase makes little contribution to inhibition by adrenalin, is at variance with the teachings of the major textbooks of biochemistry.