The recruitment of AMP-activated protein kinase to glycogen is regulated by autophosphorylation [Bioenergetics]

March 19th, 2015 by Oligschlaeger, Y., Miglianico, M., Chanda, D., Scholz, R., Thali, R. F., Tuerk, R., Stapleton, D. I., Gooley, P. R., Neumann, D.

The mammalian AMP-activated protein kinase (AMPK) is an obligatory αβγ heterotrimeric complex carrying a carbohydrate-binding module (CBM) in the β-subunit (AMPKβ) capable of attaching AMPK to glycogen. Nonetheless, AMPK localizes at many different cellular compartments, implying the existence of mechanisms that prevent AMPK from glycogen binding. Cell-free carbohydrate binding assays revealed that AMPK autophosphorylation abolished its carbohydrate-binding capacity. X-ray structural data of the CBM displays the central positioning of threonine-148 residue (T148) within the binding pocket. Substitution of T148 for a phospho-mimicking aspartate (T148D) prevents AMPK from binding to carbohydrate. Overexpression of isolated CBM or β1-containing AMPK in cellular models revealed that wild-type (WT) localizes to glycogen particles, whereas T148D shows a diffuse pattern. Pharmacological AMPK activation and glycogen degradation by glucose deprivation but not Forskolin enhanced cellular T148 phosphorylation. Cellular glycogen content was higher if pharmacological AMPK activation was combined with overexpression of T148D mutant relative to WT-AMPK. In summary, these data show that glycogen-binding capacity of AMPKβ is regulated by T148 autophosphorylation with likely implications in regulation of glycogen turnover. The findings further raise the possibility of regulated carbohydrate-binding function in a wider variety of CBM-containing proteins.