All glycogen synthesis genes were successfully cloned into BioBricks.
Muscle can't mobilize fat as quickly as it can glycogen.
Active cAMP-dependent protein kinase phosphorylates glycogen phosphorylase kinase (phosphorylase kinase), converting it from the inactive "b" for to the active "a" form. Active cAMP-dependent protein kinase phosphorylates the inhibitor 1 protein, activating it to bind to and inhibit protein phosphatase 1, and also phosphorylates protein phosphatase 1 directly, causing its inhibition. As a result the activating phosphate on phosphorylase kinase is not removed by protein phosphatase 1 and phosphorylase kinase remains active as long a glucagon or epinephrine signaling continues.
Glycogen branching enzyme
This invention relates to glycogen biosynthesis enzymes in plants. In particular, this invention is directed to plant cells having a DNA sequence encoding a glycogen biosynthesis enzyme integrated in its genome as the result of genetic engineering. Cells containing a DNA or RNA (mRNA) sequence encoding the enzyme as well as cells containing the enzyme are also provided. Plants and, more particularly, plant parts may also be obtained which contain glycogen biosynthesis enzyme sequences and/or containing such glycogen biosynthesis enzymes.
Glycogen (or Maltodextrin) + Pi
Calcium/calmodulin associates as a subunit with several enzymes and modifies their activities. It binds to inactive phosphorylase kinase, partially activating it — the fully active enzyme is both bound to the calcium/calmodulin and phosphorylated. Phosphorylase kinase phosphorylates glycogen phosphorylase b, thereby activating glycogen degradation. Calcium/calmodulin is an activator of one of the glycogen synthase kinases (calcium/calmodulin synthase kinase). Protein kinase C, calcium/calmodulin synthase kinase, and phosphorylase kinase all phosphorylate glycogen synthase, inhibiting it, and therefore, glycogen synthesis.
Glycogen (or Maltodextrin) + α-glc-1-P
In the next step, and were fused in pSB1C3 via Gibson Assembly and CPEC. We decided to put at the first position within the reading frame. The amount of ADP-glucose created by GlgC determines the activity of the following two enzymes and can therefore be considered the bottleneck of glycogen synthesis. We obtained with a point mutation in the sequence of , changing one alanine to a similiarly aliphatic aminoacid, valine. We tried to correct the substition via QuikChange and new Gibson Assembly/CPEC. Despite multiple attempts, we always received the same point mutation again. When we analysed the tertiary structure of GlgB, we assessed that the mutation was not positioned in the active site and should therefore not impede its function a lot. Due to time reasons we decided to proceed with the mutated construct which was subsequently cloned into pSB1A30 in order to confirm gene expression by .