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Powered and Designed by Full Version Forever. Characterization of the binding of glycogenin to free and glycogen-stabilized enzymes. Addition of purified glycogenin to an enzyme solution containing an AMP-sepharose column increased the interaction of the enzyme to the column. Consequently, the affinity of the enzyme to the column was increased. Even though the enzyme was adsorbed to the column to a greater extent in the presence of glycogenin, there was no increase in the specific activity of the enzyme, suggesting that binding of glycogenin to the enzyme prevented it from interacting with the column at a lower specific activity.

The mechanism s of specific interaction of glycogenin with the enzymes was studied by incubating the enzyme and glycogenin mixture at different times. At each time point, the AMP-sepharose column with the bound enzyme was washed with buffer, treated with 1 M AMP, and the eluates were assayed for specific activity.

The result showed that the binding of glycogenin to the enzyme diminished with time and at each time interval, there was a net increase in the specific activity of the enzyme, suggesting that the binding of glycogenin to the enzyme resulted in reduction of the heat lability of the enzyme. The presence of phosphorylase in the assay mixture exerted no effect on the effect of glycogenin on the activity of the enzyme.

The effect of glucosephosphate G6P on the interaction between glycogenin and the enzyme was also studied by incubating the enzyme and glycogenin mixture at different times. At each time point, G6P was added to the mixture, and the interaction of the enzyme with the AMP-sepharose column was assayed. The results indicated that G6P did not affect the interaction of the enzyme with the AMP-sepharose column. The results obtained in this study are interpreted to show that glycogenin in a free state interacts with the enzyme, forming a ternary complex in which the glycogenin-enzyme complex is stabilized by interaction with AMP-sepharose.

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