Crystallization of the glycogen-binding domain of the AMP-activated protein kinase beta subunit and preliminary X-ray analysis.
AMP-activated protein kinase (AMPK) is an intracellular energy sensor that regulates metabolism in response to energy demand and supply by adjusting the ATP-generating and ATP-consuming pathways. AMPK potentially plays a critical role in diabetes and obesity as it is known to be activated by metforin and rosiglitazone, drugs used for the treatment of type II diabetes. AMPK is a heterotrimer composed of a catalytic alpha subunit and two regulatory subunits, beta and gamma. Mutations in the gamma subunit are known to cause glycogen accumulation, leading to cardiac arrhythmias. Recently, a functional glycogen-binding domain (GBD) has been identified in the beta subunit. Here, the crystallization of GBD in the presence of beta-cyclodextrin is reported together with preliminary X-ray data analysis allowing the determination of the structure by single isomorphous replacement and threefold averaging using in-house X-ray data collected from a selenomethionine-substituted protein.
Polekhina
Galina
G
St Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia. gpolekhina@svi.edu.au
Feil
Susanne C
SC
Gupta
Abhilasha
A
O'Donnell
Paul
P
Stapleton
David
D
Parker
Michael W
MW
D055372:AMP-Activated Protein Kinases; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D017931:DNA Primers; D006003:Glycogen / Q000378:metabolism*; D009097:Multienzyme Complexes; D010446:Peptide Fragments / Q000737:chemistry; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D021122:Protein Subunits / Q000737:chemistry; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012645:Selenomethionine; D014961:X-Ray Diffraction
D017931:DNA Primers; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D021122:Protein Subunits; D011994:Recombinant Proteins; D006003:Glycogen; D012645:Selenomethionine; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases