The amino-terminal tail of glycogen phosphorylase is a switch for controlling phosphorylase conformation, activation,...

Glycogen phosphorylase is a muscle enzyme which metabolizes glycogen, producing glucose-1-phosphate, which can be used for the production of ATP. Phosphorylase activity is regulated by phosphorylation/dephosphorylation, and by the allosteric binding of numerous effectors. In this work, we have studied 10 site-directed mutants of glycogen phosphorylase (GP) in its amino-terminal regulatory region to characterize any changes that the mutations may have made on its structure or function. All of the GP mutants had normal levels of activity in the presence of the allosteric activator AMP. Some of the mutants were observed to have altered AMP-binding characteristics, however. R16A and R16E were activated at very low AMP concentration and crystallized at low temperature, like the phosphorylated form of GP, phosphorylase a, and unlike the dephospho-form, phosphorylase b. This indicates that even without phosphorylation, the structures of these mutants are more like phosphorylase a than phosphorylase b. These mutants were also very poorly phosphorylated in the presence of the inhibitor glucose, while phosphorylase b was phosphorylated normally with this inhibitor present. In contrast to R16A and R16E, four other mutants behaved like phosphorylase b after phosphorylation. R69E was only partially activated by phosphorylation, and I13G, R43E, and R43E/R69E were completely inactive after phosphorylation. We propose a model for the many functions of the amino terminus to explain the many varied effects of these mutations.